Plant disease control composition and plant disease control method

ABSTRACT

The present invention, which addresses the problem of providing a composition that has an excellent effect of controlling plant disease, provides a plant disease control composition that includes the  Bacillus  strain APM-1 (New strain of  Bacillus , APM-1) which is deposited under ATCC Accession No. PTA-4838 and at least one compound selected from the group consisting of ethaboxam, tolclofos-methyl, metalaxyl, fludioxonil, and a carboxamide compound represented by formula (1).

TECHNICAL FIELD

The present invention relates to a composition for controlling plantdiseases and a method for controlling plant diseases.

BACKGROUND ART

New strain of Bacillus, APM-1 (deposited under ATCC Accession No.PTA-4838), has been known as an active ingredient of compositions forcontrolling plant diseases and disclosed, for example, in PatentDocument 1. Also, ethaboxam, tolclofos-methyl, metalaxyl, fludioxonil,as well as a carboxamide compound represented by the formula (1)(hereinafter referred to “the carboxamide compound (1)”):

were known as an active ingredient of compositions for controlling plantdiseases and disclosed, for example, in Non-Patent Document 1 and PatentDocument 2. There is need for a material which is still more effectivefor controlling plant diseases.

PRIOR ART DOCUMENTS Patent Documents

-   Patent Document 1: WO 2003/055303-   Patent Document 2: WO 2011/162397

Non-Patent Document

-   Non-Patent Document 1: The Pesticide Manual-16th edition (BCPC,    ISBN: 978-1-901396-86-7)

DISCLOSURE OF THE INVENTION Problems to be Solved by the Invention

Damage from plant disease is a cause of considerable loss of cropproduction, and there is a need to control such plant disease moreeffectively. Thus, it is an object of the present invention to provide acomposition having an excellent controlling effect against plantdiseases.

Means for Solving the Problems

The present inventors intensively studied to achieve the above objectand have found that a composition comprising Bacillus strain APM-1 whichhas been deposited under ATCC Accession No. PTA-4838 (New strain ofBacillus, APM-1) and one or more compounds selected from the groupconsisting of ethaboxam, tolclofos-methyl, metalaxyl, fludioxonil andthe carboxamide compound (1) has an excellent controlling effect againstplant diseases.

Thus, the present invention includes the following [1] to [6].

[1] A composition for control plant diseases comprising Bacillus strainAPM-1 (New strain of Bacillus, APM-1) deposited under ATCC Accession No.PTA-4838 and one or more compounds selected from the group consisting ofethaboxam, tolclofos-methyl, metalaxyl, fludioxonil and a carboxamidecompound of the formula (1):

[2] The composition according to [1] comprising one or more compoundsselected from the group consisting of ethaboxam, tolclofos-methyl,metalaxyl, fludioxonil and a carboxamide compound of the formula (1):

in an amount of 10⁻¹⁰ to 1.5×10⁷ g per 10¹⁰ cfu of Bacillus APM-1.[3] A plant seed or a vegetative propagation organ comprising Bacillusstrain APM-1 (New strain of Bacillus, APM-1) deposited under ATCCAccession No. PTA-4838 and one or more compounds selected from the groupconsisting of ethaboxam, tolclofos-methyl, metalaxyl, fludioxonil and acarboxamide compound of the formula (1):

[4] The plant seed or vegetative propagation organ according to [3]comprising 10⁴ to 10¹⁴ cfu of Bacillus strain APM-1 and 0.000001 to 15 gof one or more compounds selected from the group consisting ofethaboxam, tolclofos-methyl, metalaxyl, fludioxonil and a carboxamidecompound of the formula (1):

per 1 kg of the seed or vegetative propagation organ.[5] A method for controlling plant diseases, comprising a step ofapplying Bacillus strain APM-1 (New strain of Bacillus, APM-1) depositedunder ATCC Accession No. PTA-4838 and one or more compounds selectedfrom the group consisting of ethaboxam, tolclofos-methyl, metalaxyl,fludioxonil and a carboxamide compound of the formula (1):

to a plant or a plant cultivation site.[6] The method for controlling plant diseases according to [5] whereinthe plant is a genetically modified plant.

Effect of Invention

The present invention provides an excellent composition for protectingseeds or vegetative propagation organs and plants grown therefrom fromplant diseases.

DESCRIPTION OF EMBODIMENTS

The composition for controlling plant diseases of the present invention(hereinafter referred to as “the present composition”) contains Bacillusstrain APM-1 (New strain of Bacillus, APM-1) deposited under ATCCAccession No. PTA-4838 (hereinafter referred to “the present bacterialstrain”) and one or more compounds selected from the group consisting ofethaboxam, tolclofos-methyl, metalaxyl, fludioxonil and the carboxamidecompound (1) (hereinafter referred to as “the present compound”).

The present bacterial strain has been disclosed in WO 2003/055303 anddeposited under the name “New strain of Bacillus, APM-1” under ATCCAccession No. PTA-4838 at ATCC (American Type Culture Collection). WO2003/055303 describes that the strain is most similar to Bucillusamyloliquefaciens. The present bacterial strain is available from ATCCand can be cultured by a known procedure. The culture may be used as itis or may be separated and concentrated using a conventional industrialtechnique, such as, not limited to, membrane separation, centrifugalseparation, or filtration separation. The fraction of the presentbacterial strain thus obtained may be used directly as it containscertain water in the present composition, or if necessary, a driedproduct obtained by a dry method, such as freeze-dry or spray drying,may be used as the present bacterial strain.

In the present composition for controlling plant diseases, the presentcompound to be used in combination with the present bacterial strain isone or more selected from the groups consisting of ethaboxam,tolclofos-methyl, metalaxyl, fludioxonil and the carboxamide compound(1).

Ethaboxam is a known compound and has been described, e.g., on page 426in “The Pesticide Manual-16th edition (Published by BCPC): ISBN978-1-901396-36-7”. Ethaboxam can be obtained from a commerciallyavailable formulation or produced by a known method.

Tolclofos-methyl is a known compound and has been described, e.g., onpage 1125 in “The Pesticide Manual-16th edition (Published by BCPC):ISBN 978-1-901396-36-7”. Tolclofos-methyl can be obtained from acommercially available formulation or produced by a known method.

Metalaxyl is a known compound and has been described, e.g., on page 733in “The Pesticide Manual-16th edition (Published by BCPC): ISBN978-1-901396-36-7”. Tolclofos-methyl can be obtained from a commerciallyavailable formulation or produced by a known method.

Fludioxonil is a known compound and has been described, e.g., on page512 in “The Pesticide Manual-16th edition (Published by BCPC): ISBN978-1-901396-36-7”. Tolclofos-methyl can be obtained from a commerciallyavailable formulation or produced by a known method.

The carboxamide compound (1) is a known compound and has been described,e.g., in the Patent Document 2. The carboxamide compound may be racemateor enantiomer, or a mixture of optional ratios of R-enantiomer andS-enantiomer and can be prepared by a known method.

The present composition can be prepared typically by mixing the presentbacterial strain and the present compound, respectively, with a solidcarrier or a liquid carrier, with addition of a surfactant or otherauxiliary agents for formulation if necessary, followed by combining thepresent bacterial strain formulation and the compound formulation thusobtained. Alternatively, the present composition can be prepared bymixing the present bacterial strain with the present compound inadvance, adding a solid carrier or a liquid carrier, with addition of asurfactant or other auxiliary agents for formulation if necessary,followed by formulating into a single formulation.

Examples of the solid carrier include mineral fine powders, such askaolin clay, pyrophyllite clay, bentonite, montmorillonite, diatomaceousearth, synthetic hydrous silicon oxide, acidic clay, talc, clay,ceramic, quartz, sericite, vermiculite, pearlite, Oya stone, anthracite,limestone, coalite, and zeolite, inorganic compounds, such as sodiumchloride, carbonate, sulfate, nitrate, and urea, organic fine powders,such as rice hulls, bran, wheat flour, and peat moss. Examples of theliquid carrier include water, vegetable oil, animal oil, and mineraloil. Examples of the auxiliary substance for formulation includeanti-freezing agents, such as ethylene glycol, and propylene glycol, andthickening agents, such as carboxymethyl cellulose, and xanthan gum.

The present composition may contain the present bacterial strain in aneffective amount, for example, at least 10⁴ cfu/g, typically 10⁴ to 10¹³cfu/g, and preferably 10⁷ to 10¹² cfu/g of the present composition.

The present composition may contain the present compound in an effectiveamount, for example, typically 0.0001 to 0.90 g, preferably 0.001 to0.80 g, per 1 g of the present composition.

The present composition typically contains 10⁻¹⁰ to 1.5×10⁷ g,preferably 10⁻⁷ to 10⁵ g, more preferably 10⁻⁵ to 10² g of the presentcompound per 10¹⁰ cfu of the present bacterial strain.

The term “effective amount” as used herein refers to an amount of thepresent bacterial strain and the present compound that is able to exertthe controlling effect against plant diseases.

The method of the invention for controlling plant diseases (hereinafterreferred to as “the present controlling method”) comprises a step ofapplying the present bacterial strain and one or more of the presentcompounds to a plant or a plant cultivation site.

In the present controlling method, the present bacterial strain and thepresent compound to be used are typically those which have beenformulated and may be applied as separate formulations or as a presentcomposition. The separate formulations may be applied simultaneously orindependently.

In the present controlling method, the present bacterial strain and thepresent compound are applied in an effective amount.

In the present invention, examples of the cultivation site of the plantinclude paddy field, cultivated field, tea field, fruit orchard,non-agricultural land, seedling tray and nursery box, nursery soil andnursery mat, water culture medium in hydroponic farm, and the like. Theplant disease may have already or not yet occurred in a cultivation siteof plant or a place of disease occurrence.

In the present controlling method, examples of the method for treatingthe present bacterial strain and the present compound include foliagetreatment, soil treatment, root treatment, seed treatment and vegetativepropagation organ treatment.

Examples of the foliage treatment include treatment of the surface ofthe cultivated plant with spraying onto the foliage and stem.

Examples of the root treatment include immersing whole plant or a rootof the plant in a solution containing the present bacterial strain andthe present compound, as well as attaching a solid preparationcontaining the present bacterial strain, the present compound and asolid carrier to a root of the plant.

Examples of the soil treatment include soil broadcast, soilincorporation and chemical irrigation to soil.

Examples of the seed treatment and vegetative propagation organtreatment include applying seed treatment or vegetative propagationtreatment using the present composition, specifically, such as spraytreatment wherein a suspension of the present composition is sprayedonto the surface of the seed or the vegetative propagation organ, wetpowder coating treatment wherein the present composition in a form ofwettable powder is coated onto moist seed or vegetative propagationorgan, smearing treatment wherein a liquid of the present compositionprepared from wettable powder, emulsifiable concentrate or flowableformulation of the present composition, with addition of water ifnecessary, is applied onto seed or vegetative propagation organ,immersion treatment wherein seeds or vegetative propagation organs areimmersed in a liquid containing the present composition for a certainperiod of time, and film coating treatment and pellet coating treatmentof seeds with the present composition.

In the present invention, the simply described “plant” encompasses inits meaning “a seed of the plant” and “a vegetative propagation organ ofthe plant”.

The term “vegetative propagation organ” as used herein means a part ofroot, stem, leaf or the like of the plant having the ability to growwhen it is separated from the body and placed on soil, such as flowerbulb, potato tuberous root, stem tuber, scaly bulb, corm, rhizophore,and strawberry runner.

In the present controlling method, the amount of the present bacterialstrain and the present compound in the treatment varies depending on thekind of plant to be treated, the kind of plant disease to be targeted,and the occurrence frequency, the formulation form, the treatmentperiod, the treatment method, the place to be treated, the weathercondition or the like, and when a stem and a leaf of the plant or a soilwhere the plant grows is treated, the amount of the present bacterialstrain for the treatment is usually 10⁵ to 10¹⁹ cfu, preferably 10⁷ to10¹⁷ cfu, per 1 ha, and the amount of the present compound for thetreatment is usually 10 to 5000 g, preferably 20 to 2000 g, per 1 ha.The composition in a form of wettable powder, water dispersible granulesor the like may be used by diluting with water so that the concentrationof the present bacterial strain is usually 10³ to 10¹² cfu/L and thatthe concentration of the present compound is usually 0.0005 to 1% byweight. The composition in a form of dustable powder or granules may beused as it is.

In the seed treatment or vegetative propagation organ treatment, theamount of the present bacterial strain is usually 10⁴ to 10¹⁴ cfu,preferably 10⁶ to 10¹³ cfu per 1 kg of the seed or vegetativepropagation organ, and the amount of the present compound is usually0.000001 to 15 g, preferably 0.0001 to 10 g, per 1 kg of the of seed orvegetative propagation organ.

The weight of the seed or vegetative propagation organ means the weightthereof when treating with the present bacterial strain and the presentcompound or other agricultural chemicals before seeding or burying ofthe same.

By treating the seed or vegetative propagation organ as described above,a seed or vegetative propagation organ comprising the present bacterialstrain and one or more compounds of the invention can be obtained. Anadjuvant may be admixed if necessary during the seed treatment orvegetative propagation organs treatment.

Examples of the plant to which the present invention is applicableinclude the followings.

Agricultural crops: cereal crops, such as corn, wheat, barley, rye, oat,sorghum; pseudocereals, such as buckwheat; pulses, such as soybean,peanut; cotton; sugar beet; rice; oilseed rape; sunflower; sugar cane;tobacco; hop.

Vegetables: solanaceous crops (eggplant, tomato, potato, chili pepper,green pepper, etc.), cucurbitaceae crops (cucumber, pumpkin, zucchini,watermelon, melon, orienta melon, etc.), cruciferous vegetables (radish,turnip, horseradish, kohlrabi, chinese cabbage, cabbage, mustard,broccoli, cauliflower, etc.), asteraceae vegetables (burdock, garlandchrysanthemum, artichoke, lettuce, etc.), liliaceae vegetables (greenonion, onion, garlic, asparagus, etc.), umbelliferae vegetables (carrot,parsley, celery, parsnip, etc.), chenopodiaceae vegetables (spinach,chard, etc.), labiatae vegetables (perilla, mint, basil, etc.),leguminous crops (pea, kidney bean, adzuki bean, broad bean, chickpea,etc.), strawberry, sweet potato, yam, taro, konjac, ginger, okra.

Fruit trees: pome fruits (apple, Japanese pear, common pear, Chinesequince, quince, etc.), stone fruits (peach, plum, nectarine, Japaneseplum, cherry, apricot, prune, etc.), citrus fruits (Satsuma mandarin,orange, lemon, lime, grapefruit, etc.), nuts (chestnut, walnut, hazelnut, almond, pistachio, cashew nut, macadamia nut, etc.), berries(blueberry, cranberry, blackberry, raspberry, etc.), grape, Japanesepersimmon, olive, loquat, banana, coffee, date palm, coconut palm, oilpalm.

Trees other than fruit trees: tea, mulberry, flowering trees (azalea,camellia, hydrangea, sasanqua, Japanese star anise, cherry, tulip tree,crape myrtle, orange osmanthus, etc.), street trees (ash tree, birch,dogwood, eucalyptus, ginkgo, lilac, maple tree, oak, poplar, cercis,Chinese sweet gum, plane tree, zelkova, Japanese arborvitae, fir tree,Japanese hemlock, needle juniper, pine, spruce, yew, elm, horsechestnut, etc.), coral tree, podocarpus, cedar, Japanese cypress,croton, Japanese spindle tree, Japanese photinia.

Grasses: zoysia (zoysiagrass, Zoysia matrella, etc.), bermuda grasses(Cynodon dactylon, etc.), bent grasses (Agrostis alba, creeping bentgrass, hiland bent, etc.), blueglasses (meadow grass, bird grass, etc.),fescue (tall fescue, chewings fescue, creeping red fescue, etc.),ryegrasses (darnel, rye grass, etc.), orchard grass, timothy grass.

Others: flowers (rose, carnation, chrysanthemum, prairie gentian,gypsophila, gerbera, marigold, salvia, petunia, verbena, tulip, aster,gentian, lily, pansy, cyclamen, orchid, convallaria, lavender, stock,ornamental cabbage, primula, poinsettia, gladiolus, cattleya, daisy,cymbidium, begonia, etc.), bio-fuel plants (Jatropha, safflower,camelina, switchgrass, Miscanthus, reed canary grass, giant reed, kenaf,cassava, willow, etc.), ornamental plants.

The present invention is preferably applied to cereal crops or millets.The present invention is more preferably applied to corn, wheat,sorghum, and soybean.

In the present invention, the variety of plant is not limited so long asit is commonly cultivated. The plants of such varieties include plantswhich have been conferred with one or more useful trait by a classicalbreeding technique or a genetic engineering technique (geneticallymodified plant) as well as stack varieties obtained by crossing suchgenetically modified plants.

Such useful characters include tolerance to herbicide, resistance toplant disease, disease resistance, stress tolerance, and improvedquality of crops such as modified fatty acid residue composition of oilsand fats.

Examples of the genetically modified plant include those listed in thegenetically modified crop registration database (GM APPROVAL DATABASE)in the electronic information site (http://www.isaaa.org/) of theINTERNATINAL SERVICE for the ACQUISITION of AGRI-BIOTECH APPLICATIONS(ISAAA). More specifically, the plant may be a plant which has beenconferred with an environmental stress tolerance, a disease resistance,a herbicide tolerance, a pest resistance or the like, or a plant whereinits trait has been modified with respect to growth and yield, quality ofproduct, sterility or the like, by genetic recombination technology.

Examples of the plant conferred with a herbicide tolerance by generecombination technology include genetically modified plants conferredwith a tolerance to protoporphyrinogen oxidase (herein after referred toas PPO) herbicides such as flumioxazin; 4-hydroxyphenyl pyruvic aciddioxygenase (hereinafter abbreviated as HPPD) inhibitors such asisoxaflutole, mesotrione; acetolactate synthase (hereinafter referred toas ALS) inhibitors such as imazethapyr, thifensulfuron methyl;5-enolpyruvylshikimate-3-phosphate synthase (hereinafter referred to asEPSP) inhibitors such as glyphosate; glutamine synthetase inhibitorssuch as glufosinate; auxin herbicides such as 2,4-D, dicamba; andherbicides such as bromoxynil.

Examples of the plant conferred with a herbicide tolerance by generecombination technology include glyhosate-tolerant genetically modifiedplants which have been introduced with one or more gene selected fromglyphosate tolerant EPSPS gene (CP4 epsps) from Agrobacteriumtumefaciens strain CP4; glyphosate metabolizing enzyme gene (gat4601,gat6421) which is a gene of glyphosate metabolizing enzyme (glyphosateN-acetyl transferase) from Bacillus (Bacillus licheniformis) modified bygene shuffling to enhance the metabolic activity; glyphosatemetabolizing enzyme (glyphosate oxidase gene, goxv247) from Ochrobactrum(Ochrobactrum anthropi strain LBAA), or EPSPS gene havingglyphosate-tolerant mutation (mepsps, 2mepsps) from corn. There areglyphosate-tolerant genetically modified varieties with respect toplants such as corn (Zea mays L.), soybean (Glycine max L.), cotton(Gossypium hirsutum L.), sugar beet (Beta vulgaris), canola (Brassicanapes, Brassica rapa), alfalfa (Medicago sativa), potato (Solanumtuberrosum L), wheat (Triticum aestivum), and creeping bent grass(Agrostis stolonifera).

Some glyphosate-tolerant genetically modified plants are commerciallyavailable. For example, a genetically modified plant expressingglyphosate-tolerant EPSPS from Agrobacterium has been marketed under thetrade name such as Roundup Ready®, a genetically modified plantexpressing glyphosate metabolizing enzyme from Bacillus with enhancedmetabolic activity by gene shuffling has been marketed under the tradename such as Optimum® GAT®, Optimum® Gly canola, and a geneticallymodified plant expressing EPSPS gene having glyphosate-tolerant mutationhas been marketed under the trade name GlyTol®.

Examples of plants conferred with herbicide-tolerance by generecombination technology include glufosinate-tolerant geneticallymodified plants which have been introduced with phosphinothricinN-acetyltransferase (PAT) gene (bar) of the glufosinate metabolizingenzyme from Streptomyces (Streptomyces hygroscopicus), phosphinothricinN-acetyltransferase gene (pat) of the glufosinate metabolizing enzymefrom Streptomyces (Streptomyces viridochromogenes), a synthesized patgene, or the like. There are glufosinate-tolerant genetically modifiedvarieties with respect to plants such as corn, soybean, cotton, canola,rice (Oryza sativa L.), sugar beet, and cichory (Cichori intybus).

Some glufosinate-tolerant genetically modified plants are commerciallyavailable. A genetically modified plant expressing glufosinatemetabolizing enzyme (bar, pat) from Streptomyces has been marked under atrade name including LibertyLink®.

Examples of herbicide-tolerant genetically modified plants includegenetically modified plants which have been introduced with the gene(bxn) of nitrilase, which is a bromoxynil-metabolizing enzyme fromKlebsiella (Klebsiella pneumoniae subsp. Ozaenae). Bromoxynil-tolerantgenetically modified varieties have been produced for plants such ascanola, cotton, tobacco (Nicotiana tabacum L.) and have been markedunder a trade name including Navigator® canola, or BXN®.

Examples of herbicide-tolerant genetically modified plants includegenetically modified carnation (Dianthus caryophyllus) which has beenintroduced with ALS herbicide-tolerant ALS gene (SurB, S4-HrA) fromtobacco as a selectable marker. Also, a genetically modified larvae(Linum usitatissumum L.) which has been introduced with ALSherbicide-tolerant ALS gene from Arabidopsis (Arabidopsis thaliana) hasbeen developed under the trade name CDC Triffid Flax. Also, agenetically modified soybean which has been introduced with ALSherbicide-torelant ALS gene (csr1-2) from Arabidopsis has been developedunder the trade name Cultivance®. Furthermore, there aresulfonylurea/imidazolinone herbicide-tolerant genetically modified cornwhich has been introduced with ALS herbicide-tolerant ALS gene (zm-hra)from corn, and sulfonylurea herbicide-tolerant genetically modifiedsoybean which has been introduced with ALS herbicide-tolerant ALS gene(gm-hra) from soybean.

Examples of plants conferred with herbicide-tolerance by generecombination technology include isoxaflutole-tolerant geneticallymodified soybean which has been introduced with HPPD herbicide-tolerantHPPD gene (hppdPFW 336) from Pseudomonas (Pseudomonas fluorescens strainA32) and mesotrione-tolerant genetically modified soybean which has beenintroduced with HPPD gene (avhppd-03) from oats (Avena sativa).

Examples of plants conferred with herbicide-tolerance by generecombination technology include 2,4-D-tolerant genetically modifiedcorns, genetically modified soybeans, genetically modified cottons whichhave been introduced with gene (aad-1) of 2,4-D metabolizing enzymearyloxyalkanoate dioxygenase from Sphingobium (Sphingobiumherbicidovorans) or with gene (aad-12) of 2,4-D metabolizing enzymearyloxyalkanoate dioxygenase from Delftia (Delftia acidovorans). Some ofthem are developed under the trade names such as Enlist® Maize, Enlist®Soybean. Also, there are dicamba-tolerant genetically modified soybeansand cottons which have been introduced with gene (dmo) of dicambamonooxygenase, which is dicamba metabolizing enzyme fromStenotrophomonas (Stenotrophomonas maltophilia strain DI-6).

Examples of genetically modified plant tolerant to two or moreherbicides include geneticaly modified cotton and genetically modifiedcorn, which are tolerant to both glyphosate and glufosinate, andmarketed under the trade name such as GlyTol® LibertyLink®, RoundupReady® LibertyLink® Maize. Also, there are a genetically modifiedsoybean tolerant to both glufosinate and 2,4-D and developed under thetrade name Enlist® Soybean, and a genetically modified cotton tolerantto both glufosinate and 2,4-D. A genetically modified soybean tolerantto both glyphosate and dicamba has been developed under the trade nameGenuity®) Roundup Ready® 2 Xtend®. Genetically modified corn and soybeanresistant to both glyphosate and ALS inhibitors have been developedunder the trade name Optimum® GAT®. In addition, a genetically modifiedcotton tolerant to both glufosinate and dicamba, a genetically modifiedcorn tolerant to both glyphosate and 2,4-D, a genetically modifiedsoybean tolerant to both glyphosate and HPPD herbicide have also beendeveloped. Furthermore, a genetically modified soybean tolerant to threeherbicides glyphosate, glufosinate and 2,4-D has been developed.

Examples of the plant conferred with a pest resistance by generecombination technology include plants conferred with resistance tolepidopteran insects, coccinella insects, multipter insects, nematodesand the like.

Examples of the plant conferred with a pest resistance to lepidopteraninsects by genetic recombination technology include genetically modifiedplants such as soybean, cotton, rice, poplar (Populus sp.), and tomato(Lycopersicon esculentum), and eggplant (Solanum melongena), which havebeen introduced with a gene encoding delta-endotoxin, which is aninsecticidal protein derived from a soil bacterium Bacillusthuringiensis bacteria (hereinafter referred to as Bt bacteria).Examples of the delta-endotoxin that confers a pest resistance tolepidopteran insects include Cry1A, Cry1Ab, modified Cry1Ab (truncatedCry1Ab), Cry1Ac, Cry1Ab-Ac (hybrid protein of Cry1Ab and Cry1Ac), Cry1C,Cry1F, Cry1Fa2 (modified cry1F), moCry1F (modified Cry1F), Cry1A. 105(hybrid protein of Cry1Ab, Cry1Ac and Cry1F), Cry2Ab2, Cry2Ae, Cry9C,Vip3A, Vip3Aa20, and the like.

Examples of the plant conferred with a pest resistance to coccinellainsects by genetic recombination technology include genetically modifiedplants such as corn, potato, which have been introduced with a geneencoding delta-endotoxin, which is an insecticidal protein derived froma soil bacterium Bt bacteria. Examples of the delta-endotoxin thatconfers a pest resistance to coccinella insects include Cry3A, mCry3A(modified Cry3A), Cry3Bb1, Cry34Ab1, and Cry35Ab1.

Examples of the plant conferred with a pest resistance to multipterinsects by genetic recombination technology include genetically modifiedcorn, which has been introduced with a synthetic gene encoding a hybridprotein eCry3.1Ab, which is a hybrid protein of Cry3A and Cry1Ab derivedfrom soil bacteria Bt bacteria, a genetically modified cotton, which hasbeen introduced with a gene encoding trypsin inhibitor CpTI fromblack-eyed pea (Vigna unguiculata), a genetically modified poplar, whichhas been introduced with a gene encoding API, which is a proteaseinhibitor protein A from arrowhead (Sagittaria sagittifolia).

Examples of the insecticidal protein that confers a pest resistance tothe plants include hybrid proteins, truncated proteins, and modifiedproteins of the insecticidal proteins described above. The hybridproteins are produced by combining different domains of multipleinsecticidal proteins using a common recombination technology, andCry1Ab-Ac and Cry1A.105 are known.

Examples of the truncated proteins include Cry1Ab lacking the amino acidsequence partially. Examples of the modified proteins include proteinsin which one or more amino acids of natural delta-endotoxin have beensubstituted, such as Cry1Fa2, moCry1F, mCry3A.

Examples of other insecticidal proteins that confer insect resistance toplants by genetic recombination technology include insecticidal proteinsfrom Bacillus cereus or Bacillus popilliae, the insecticidal proteinsVip 1, Vip 2, Vip 3 of Bt bacteria, insecticidal proteins from nematode,toxin produced by an animal such as scorpotoxin, spider toxin, bee venomor insect-specific neurotoxin, toxins of filamentous fungi, plantlectin, agglutinin, protease inhibitor such as trypsin inhibitor, serineprotease inhibitor, patatin, cystatin, papain inhibitor, ribosomeinactivating protein (RIP) such as ricin, corn-RIP, abrin, rufin,saporin, bryodin, steroid metabolizing enzymes such as 3-hydroxysteroidoxidase, ecdysteroid-UDP-glucosyltransferase, cholesterol oxidase,ecdysone inhibitor, HMG-CoA reductase, ion channel inhibitors such assodium channel inhibitor, calcium channel inhibitor, juvenile hormoneesterase, diuretic hormone receptor, stilbene synthase, bibenzylsynthase, chitinase, glucanase, and the like.

Genetically modified plants conferred with a pest resistance byintroducing one or more insecticidal protein gene are known, and some ofsuch genetically modified plants are commercially available.

Examples of commercially available genetically modified cotton conferredwith a pest resistance include Bollgard® cotton expressing theinsecticidal protein Cry1Ac of Bt bacteria, Bollgard II® cottonexpressing the insecticidal proteins Cry1Ac and Cry2Ab of Bt bacteria,Bollgard III® expressing the insecticidal proteins Cry1Ac, Cry2Ab, Vip3Aof Bt bacteria, VIPCOT® expressing the insecticidal proteins Vip3A andCry1Ac of Bt bacteria, WideStrike® expressing the insecticidal proteinsCry1Ac, Cry1F of Bt bacterium.

Examples of commercially available genetically modified corn conferredwith a pest resistance include YieldGard® Rootworm RW expressing theinsecticidal protein Cry3Bb1 of Bt bacteria, YieldGard Plus® expressingthe insecticidal proteins Cry1Ab and Cry3Bb1 of Bt bacteria, YieldGard®VT Pro® expressing the insecticidal proteins Cry1A.105 and Cry2Ab2 of Btbacteria. Agrisure® RW expressing the insecticidal protein mCry3A of Btbacteria, Agrisure® Viptera expressing the insecticidal protein Vip3Aa20of Bt bacteria, Agrisure® Duracade® expressing the insecticidal proteineCry3.1Ab of Bt bacteria are also commercially available.

Examples of commercially available genetically modified potato conferredwith a pest resistance include Atlantic NewLeaf® potato, NewLeaf® RussetBurbank potato, and the like, which express the insecticidal proteinCry3A of Bt bacteria.

Examples of genetically modified plants conferred with resistance toplant diseases include kidney bean (Phaseolus vulgaris), papaya (Caricapapaya), plum (Prunus domestica), potato, squash (Cucurbita pepo), sweetpepper (Capsicum annuum), tomato, and the like, which have beenconferred with a resistance to plant viral diseases. Specific examplesof genetically modified plants conferred with a resistance to plantviral diseases include a genetically modified kidney bean which has beenintroduced with a gene that produces double-stranded RNA of areplication protein of bean golden mosaic virus, a genetically modifiedpapaya which has been introduced with a coat protein gene of papayaringspot virus, a genetically modified potato which has been introducedwith a coat protein gene of potato virus Y or replication enzyme domaingene of potato leaf roll virus, a genetically modified squash which hasbeen introduced with a coat protein gene of Cucumber mosaic⋅virus, witha coat protein gene of Watermelon mosaic virus, or with a coat proteingene of Zucchini yellow mosaic virus, a genetically modified sweetpepper and transgenic tomato which has been introduced with a coatprotein gene of Cucumber mosaic⋅virus, and the like.

A genetically modified potato conferred with a resistance to plant viraldiseases is commercially available under a trade name includingNewLeaf®.

Examples of the plant conferred with a resistance to plant disease alsoinclude plants that have been conferred with an ability to produce aselective anti-pathogenic substance using genetic recombinationtechnology. PR proteins are known as an anti-pathogenic substance (PRPs,EP392225). Such anti-pathogenic substance and genetically modifiedplants that produce the same are described in EP 392225, WO 199533818,EP 353191 and the like. Examples of the anti-pathogenic substanceinclude ion channel inhibitors such as sodium channel inhibitors,calcium channel inhibitors (KP1, KP4, KP6 toxin produced by viruses areknown), anti-pathogenic substances produced by microorganisms such asstilbene synthase, bibenzyl synthase, chitinase, glucanase, peptideantibiotics, antibiotics having heterocycles, protein factors involvedin plant disease resistance, which is referred to as plant diseaseresistance genes and described in WO 2003000906.

Examples of genetically modified plant wherein the quality of producthas been modified includes genetically modified plants having amodification in lignin production, a modification in oils or fatty acidcomponents, production of phytic acid degrading enzymes, a modificationin flower color, a modification in alpha-amylase activity, amodification in amino acids, a modification in starch or carbohydratecomponents, inhibition of acrylamide production, reduction of blackspots due to mechanical damage, anti-allergy, reduction of nicotineproduction, or retardation of aging or grain-filling.

There is a genetically modified alfalfa wherein the lignin content hasbeen lowered by RNA interference with a gene that generatesdouble-stranded RNA of S-adenosyl-L-methionine: trans-caffeoyl CoA3-methyltransferase (ccomt) gene of alfalfa related to ligninproduction.

A genetically modified canola wherein the triacylglyceride content,including lauric acid, has been increased by introducing a gene involvedin fatty acid synthesis, 12:0 ACP thioesterase gene of laurier(Umbellularia californica), has been developed under the trade nameLaurical® Canola.

A genetically modified canola wherein the degradation of endogenousphytic acid has been enhanced by introducing a gene (phyA) of 3-phytase,which is a degrading enzyme of phytic acid of plants from Aspergillusniger, has been developed under the trade name Phytaseed® Canola. Also,a genetically modified corn wherein the degradation of endogenous phyticacid has been enhanced by introducing 3-phytase gene (phyA) ofAspergillus niger has been developed.

A genetically modified carnation wherein the flower color has beencontrolled to blue by introducing a gene of dihydroflavonol-4-reductase,which is an enzyme that produces blue pigment delphinidin and itsderivative of petunia (Petunia hybrida), and aflavonoid-3′,5′-hydroxylase gene from petunia, pansy (Violawittrockiana), salvia (Salvia splendens) or carnation is known.Genetically modified carnations with flower color controlled to bluehave been developed under the trade name such as Moonldust®,Moonshadow®, Moonshade®, Moonlite®, Moonaqua®, Moonvista®, Moonique®,Moonpearl®, Moonberry Registered trademark), and Moonvelvet®. Also,genetically modified roses with flower color controlled to blue byintroducing a gene of anthocyanin-5-acyltransferase, which is an enzymethat produces blue pigment delphinidin and its derivative, from Torenia(Torenia sp.), and a flavonoid-3′,5′-hydroxylase gene from pansy havebeen developed.

A genetically modified corn wherein the production of bioethanol hasbeen increased by introducing a gene (Amy797E) of heat-resistantalpha-amylase relating to starch degradation of Thermococcales sp. havebeen developed under the trade name Enogen®.

A genetically modified corn wherein the production of lysine has beenincreased by introducing a gene (cordapA) of dihydrodipicolinatesynthase relating to the production of amino acid lysine ofCorynebacterium glutamicum has been developed under the trade nameincluding Mavera®.

A genetically modified melon and a genetically modified tomato whereinthe shelf life has been improved by introducing a gene (sam-K) ofS-adenosylmethionine hydrolase relating to ethylene production by planthormones from Escherichia coli bacteriophage T3 has been developed.Also, genetically modified tomatoes with improved shelf life byintroducing a gene that lacks a part of the ACC synthase gene, which isinvolved in the ethylene production by plant hormones, from tomato, anACC deaminase gene from Pseudomonas (Pseudomonas chlororaphis) thatdegrades the ethylene precursor ACC, a gene that generatesdouble-stranded RNA of polygalacturonase genes which degrades cell wallpectin, or ACC oxidase genes of tomato related to the production ofethylene have been developed. A genetically modified tomato withimproved shelf life by introducing a gene that produces double-strandedRNA of polygalacturonase genes of tomato has been developed under thetrade name FLAVR SAVR®.

A genetically modified potato, wherein the possibility of decompositionof starch, formation of black spots due to mechanical damage andproduction of a carcinogen (acrylamide) from heating are lowered byintroducing a gene that generates double-stranded RNA of a transcriptionfactor promoting degradation of starch derived from potato, and a genethat generates double-stranded RNA of polyphenol oxidase gene and a genethat generates double-stranded RNA of genes involved in asparagineproduction from potato, has been developed under a trade mark includingInnate®. Also, a genetically modified potato wherein the amylose contentis lowered by introducing an antisense gene of starch synthase frompotato has been developed under the trade name Amflora®.

A genetically modified rice having alleviation effect on pollinosis withimmune tolerance by introducing a gene (7crp) of altered antigenicprotein of cedar pollen has been developed.

A genetically modified soybean wherein the oleic acid content isincreased by introducing a partial gene (gm-fad2-1) of ω-6 desaturase,which is a fatty acid desaturase enzyme, of soybean to inhibit the geneexpression thereof has been developed under the trade name Plenish® orTreus®. Also, a genetically modified soybean wherein the saturated fattyacid content is lowered by introducing a gene (fatb1-A) that generates adouble-stranded RNA of acyl-acyl carrier protein-thioesterase and a gene(fad2-1A) that generates a double-stranded RNA of δ-12 desaturase hasbeen developed under the trade name Vistive Gold®. Also, a geneticallymodified soybean wherein the ω3 fatty acid content is enhanced byintroducing a δ-6 desaturase gene (Pj.D6D) of primrose and a δ-12desaturase gene (Nc.Fad3) of Neurospora crassa has been developed.

A genetically modified tobacco wherein the nicotine content is loweredby introducing an antisense gene of quinolinic acidphosphoribosyltransferase (NtQPT1) of tobacco has been developed.

A genetically modified rice, Golden rice, introduced with a phytoenesynthase gene (psy) of trumpet narcissus (Narcissus pseudonarcissus) anda carotene desaturase gene (crt1) of soil bacteria that synthesizescarotenoids (Erwinia uredovora), which allow endosperm-specificexpression to produce β-carotene in endosperm tissue, whereby a ricecontaining vitamin A is enabled to be harvested, has been developed.

Examples of the plants in which the fertile trait has been modified by agenetic recombination technique include genetically modified plantsconferred with male sterility and fertility restoration. There aregenetically modified corn and chicory conferred with male sterility byintroducing anther tapetum cell expressing a ribonuclease gene (barnase)of Bacillus (Bacillus amyloliquefaciens). There is also a geneticallymodified corn conferred with male sterility by introducing a DNA adeninemethyltransferase gene (dam) of Escherichia coli. Furthermore, there isa genetically modified corn wherein the sterility has been controlled byintroducing alpha-amylase gene (zm-aa1) of corn that confers malesterility and ms45 protein gene (ms45) of corn that confers fertilityrestoration.

There is a genetically modified canola conferred with a fertilityrestoring function by introducing anther tapetum cells expressing aribonuclease inhibitory protein gene (barstar) of Bacillus. In addition,there is a genetically modified canola wherein the sterility has beencontrolled by introducing a ribonuclease gene (barnase) of Bacillus thatconfers a male sterility and a ribonuclease inhibitory protein gene(barstar) of Bacillus that confers a fertility restoration.

Examples of the plants conferred with tolerance to environmental stressby a genetic recombination technique include genetically modified plantsconferred with tolerance to dryness. A dry tolerant corn which has beenintroduced with a cold shock protein gene (cspB) of Bacillus subtilishas been developed under the trade name Genuity® DroughtGard®. Also, drytolerant sugar cane which has been introduced with choline dehydrogenasegene (RmBetA) of alfalfa rhizobium (Rhizobium meliloti) or E. coli(Esherichia coli) has been developed.

Examples of the plants wherein a trait related to growth and yield hasbeen modified by genetic recombination technology include geneticallymodified plants having enhanced growth ability. For example, agenetically modified soybean which has been introduced with a gene ofArabidopsis encoding a transcription factor that controls circadianrhythm (bbx32) has been developed.

The plant according to the present invention can be a plant which hasbeen modified using other techniques than genetic recombinationtechnology. More specifically, it may be a plant which has beenconferred with tolerance to environmental stress, disease resistance,tolerance to herbicide, insect resistance, or the like, by classicalbreeding technique, genetic marker breeding technique, genome editingtechnique, or the like.

Examples of the plant wherein a tolerance to herbicide has beenconferred by classical breeding technique or genetic marker breedingtechnique include corn, rice, wheat, sunflower (Helianthus annuus),canola, and lentil beans (Lens culinaris), which are resistant toimidazolinone type ALS inhibiting herbicides, such as imazethapyr, andare marketed under the trade name Clearfield®. Also, there is STSsoybean, which is a soybean tolerant to sulfonylurea-based herbicide, asan example of plants which has been conferred with a resistance tosulfonyl-based ALS-inhibiting herbicides such as thifensulfuron methylby genetic marker breeding technique. Also, there is SR corn, which isresistant to sethoxydim, as an example of plants which has beenconferred with a resistance to acetyl CoA carboxylase inhibitor, such astrione oxime type herbicide, aryloxyphenoxypropionic acid typeherbicide, by genetic marker breeding technique.

Examples of the plants conferred with pest resistance by classic orgenetic marker breeding technique include a soybean having Rag 1(Resistance Aphid Gene 1) gene, which is an aphid resistant gene.Examples of the plants conferred with resistance to nematodes by theclassical breeding technique include a soybean conferred with aresistance to Cysto nematode, and a cotton conferred with a resistanceto Root Knot nematode.

Examples of the plants which has been conferred with a resistance toplant disease by classic or genetic marker breeding technique include acorn which has been conferred with a resistant to anthracnose stalk rot,a corn which has been conferred with a resistant to Gray leaf spot, acorn which has been conferred with a resistant to Goss's wilt, a cornwhich has been conferred with a resistant to Fusarium stalk rot, asoybean which has been conferred with a resistant to Asian soybean rust,a pepper which has been conferred with a resistant to Phytophthora, alettuce which has been conferred with a resistant to powdery mildew, atomato which has been conferred with a resistant to Bacterial wilt, atomato which has been conferred with a resistant to Gemini virus, and alettuce which has been conferred with a resistant to downy mildew.

As an example of the plants which have been conferred with a toleranceto dryness by classic or genetic marker breeding technique, a drytolerant corn has been developed under the trade name such as AgrisureArtesian®, Optimum AQUA Max®.

As an example of the plants conferred with a tolerance to herbicide bygenomic editing technique, a canola conferred with a tolerance tosulfonylurea herbicide by rapid breed development technology wherein amutation to confer tolerance to sulfonylurea herbicide has introducedinto ALS gene via chimera oligonucleotides of DNA and RNA.

The above plants include a variety which has been conferred with two ormore traits, such as tolerance to environmental stress, diseaseresistance, tolerance to herbicide, pest resistance, growth and yieldtraits, quality of product, and sterility, using a genetic recombinationtechnology as described above, such as a classic breeding technique, agenetic marker breeding, or a genome editing technique, as well as avariety which has been conferred with two or more traits from parents bycrossing the parents, which are genetically modified plants having sameor different characteristic. Examples of such plant include geneticallymodified plants conferred with both of tolerance to herbicide and pestresistance.

For example, as for a genetically modified plant conferred withtolerance to glyphosate and pest resistance, genetically modifiedcottons, such as Roundup Ready® Bollgard® cotton, Roundup Ready®Bollgard II® cotton, Roundup Ready® Flex® Bollgard II® cotton, Bollgard®III×Roundup Ready® Flex®, and VIPCOT® Roundup Ready Flex® Cotton, havebeen developed. Also, genetically modified soybeans have been developedunder the trade name, such as Agrisure® GT/RW, Roundup Ready® YieldGard®maize, Genuity® VT Double Pro®, Genuity® VT Triple Pro®, YieldGard®,YieldGard® CB+RW, YieldGard® VT® Rootworm® RR 2, YieldGard® RW+RR,YieldGard® VT Triple, or YieldGard® Plus with RR. Furthermore, agenetically modified soybean such as Intacta® Roundup Ready® 2 Pro hasbeen developed.

For example, as for genetically modified plants conferred with toleranceto glufosinate and pest resistance, genetically modified cottons havebeen developed under the trade name, such as Widestrike® Cotton,Twinlink® Cotton, and FiberMax® LibertyLink® Bollgard II®. Also,genetically modified corns have been developed under the trade name,such as Agrisure® CB/LL, Agrisure® CB/LL/RW, Agrisure® Viptera® 2100,Agrisure® Viptera® 3100, Bt Xtra Maize, NaturGard Knockout®, Herculex®RW, Herculex® CB, Herculex® XTRA, Starlink® Maize, and Liberty Link®YieldGard® Maize.

For example, as for genetically modified plants conferred with toleranceto glyphosate and glufosinate and pest resistance, genetically modifiedcottons have been developed under the trade name, such as Widestrike®Roundup Ready® Cotton, Widestrike® Roundup Ready Flex® Cotton,Widestrike® Cotton, Registered trademark)×Roundup Ready Flex®×VIPCOT®Cotton, and Glytol®×Twinlink®. Also, genetically modified corns havebeen developed under the trade name, such as Agrisure® GT/CB/LL,Agrisure® 3000GT, Agrisure® 3122, Agrisure® Viptera® 3110, Agrisure®Viptera 3111, Agrisure® Viptera® 3220, Agrisure® Duracade® 5122,Agrisure® Duracade® 5222, Optimum® Intrasect, Optimum® TRIsect, Optimum®Intrasect XTRA, Optimum® Intrasect Xtreme, Genuity® martStax®, PowerCore®, Herculex® I RR, Herculex® RW Roundup Ready® 2, and Herculex XTRA®RR.

For example, as for genetically modified plants conferred with toleranceto bromoxynil and pest resistance, a genetically modified cottons hasbeen developed under the trade name, such as BXN® Plus Bollgard® Cotton.

Examples of a variety conferred with two or more traits includegenetically modified plants conferred with disease resistance and pestresistance. For example, as for genetically modified plants conferredwith resistance to potato virus Y and pest resistance, geneticallymodified potatoes have been developed under the trade name, such asHi-Lite NewLeaf® Y Potato, NewLeaf® Y Russet Burbank Potato, and ShepodyNewLeaf® Y potato. As for genetically modified plants conferred withresistance to potato leaf roll virus and pest resistance, geneticallymodified potatoes have been developed under the trade name, such asNewLeaf® Plus Russet Burbank Potato.

Examples of a variety conferred with two or more traits includegenetically modified plants conferred with tolerance to herbicide andaltered product quality. For example, a genetically modified canola andgenetically modified corn, which have been conferred with tolerance toglufosinate and fertile trait have been developed under the trade name,such as InVigor® Canola and InVigor® Maize, respectively.

Examples of a variety conferred with two or more traits includegenetically modified plants conferred with a pest resistance and alteredproduct quality. For example, a genetically modified corn conferred withresistance to lepidopterous insects and a trait of enhanced lysineproduction has been developed under the trade name such as Mavera®YieldGard® Maize.

For other Examples of a variety conferred with two or more traits asmentioned above, genetically modified plants conferred with tolerance toherbicide and a trait altering fertility, genetically modified plantsconferred with tolerance to herbicide and tolerance to environmentalstress, genetically modified plants conferred with tolerance toherbicide and a trait modifying growth and yield, genetically modifiedplants conferred with tolerance to herbicide, pest resistance, and atrait modifying product quality, genetically modified plants conferredwith tolerance to herbicide, pest resistance, and tolerance toenvironmental stress, have been developed.

Examples of the plant diseases which can be controlled according to thepresent invention include the followings.

Diseases of rice: blast (Magnaporthe oryzae), brown spot (Cochliobolusmiyabeanus), sheath blight (Rhizoctonia solani), “Bakanae” disease(Gibberella fujikuroi), seedling blight (Pythium arrhenomanes, Pythiumgraminicola, Pythium spinosum, Pythium sp., Rhizopus chinensis, Rhizopusoryzae, Trichoderma viride);

Diseases of wheat: powdery mildew (Erysiphe graminis), Fusarium blight(Fusarium graminearum, F. avenaceum, F. culmorum, F. asiaticum,Microdochium nivale), rust (Puccinia striiformis, P. graminis, P.recondita, P. hordei), snow mold (Typhula sp., Micronectriella nivalis),loose smut (Ustilago tritici, U. nuda), stinking smut (Tilletia caries),eye spot (Pseudocercosporella herpotrichoides), scald (Rhynchosporiumsecalis), speckled leaf blotch (Septoria tritici), glume blotch(Leptosphaeria nodorum), net blotch (Pyrenophora teres Drechsler),yellow spot (Pyrenophora tritici-repentis), stripe (Pyrenophoragraminea), Rhizoctonia damping-off (Rhizoctonia solani), snow mold(Typhula ishikariensis, Typhula incarhata, Sclerotinia borealis,Microdochium nivale), foot lot disease (Fusarium graminearum);

Diseases of corn: smut (Ustilago maydis), brown spot (Cochliobulusheterostrophus), zonate leaf spot (Gloeocercospora sorghi), southernrust (Puccinia polysora), grey leaf spot (Cercospora zaea-maydis),Rhizoctonia danpimg-off (Rhizoctonia solani), gibberella ear rot(Fusarium moniliforme), anthracnose (Colletotrichum graminicola),seedling blight (Fusarium spp, Rhizoctonia solani);Diseases of citrus: gray mold (Botrytis cinerea), black leaf spot(Diaporthe citri), scab (Elsinoe fawcetti), fruit rot (Penicilliumdigitatum, P. italicum); brown rot (Phytophthora parasitica,Phytophthora citrophthora);Diseases of apple: Monilia leaf blight (Monilinia mali), Valsa canker(Valsa ceratosperma), powdery mildew (Podosphaera leucotricha),Alternaria blotch (Alternaria alternate apple pathotype), scab (Venturiainaequalis), anthracnose (Colletotrichum gloeosporioies, Colletotrichumacutatum), Phytophthora rot (Phytophtora cactorum); blotch (Diplocarponmali); ring rot (Botryosphaeria berengeriana);Diseases of pear: scab (Venturia nashicola, V. pirina), black spot(Alternaria alternate Japanese pear pathotype), rust (Gymnosporangiumharaeanum), Phytophthora fruit rot (Phytophthora cactorum);Diseases of peach: brown rot (Monilinia fructicola), scab (Cladosporiumcarpophilum), Phomopsis seed decay (Phomopsis sp.);Diseases of grape: anthracnose (Elsinoe ampelina), ripe rot(Colletorichum gloeosporioides, Colletotrichum acutatum), powdery mildew(Uncinula necator), rust (Phakopsora ampelopsidis), black rot(Guignardia bidwellii), downy mildew (Plasmopara viticola), gray mold(Botrytis cinerea);Diseases of persimmon: anthracnose (Gloeosporium kaki), leaf spot(Cercospora kaki, Mycosphaerella nawae);Diseases of cucumber: anthracnose (Colletotrichum orbiculare), powderymildew (Sphaerotheca fuliginea), gummy stem blight (Mycosphaerellamelonis), Fusarium wilt (Fusarium oxysporum), downy mildew(Pseudoperonospora cubensis), Phytophthora blight (Phytophthora sp.),damping-off (Pythium sp.); Rhizoctonia damping-off (Rhizoctonia solani),gray mold (Botrytis cinerea);Diseases of tomato: Early blight (Alternaria solani), Leaf mold(Cladosporium flavum), late blight (Phytophthora infestans), leaf spot(Stemphylium lycoperici), gray mold (Botrytis cinerea);Diseases of eggplant: brown spot (Phomopsis vexans), powdery mildew(Erysiphe cichoracearum), gray mold (Botrytis cinerea);Diseases of brassica vegetables: Alternaria leaf spot (Alternariajaponica), leaf spot (Cercosporella brassicae), Clubroot (Plasmodiophorabrassicae), downy mildew (Peronospora parasitica), root rot (Phomalingam);Diseases of rapeseed: Sclerotinia rot (Sclerotinia sclerotiorum),Alternaria leaf spot (Alternaria brassicae), powdery mildew (Erysiphecichoracearum), black leg (Leptosphaeria maculans), Rhizoctoniadamping-off (Rhizoctonia solani);Diseases of green onion: rust (Puccinia allii), Fusarium wilt (Fusariumoxysoporum);Diseases of onion: gray-mold neck rot (Botrytis allii), leaf blight(Botrytis squamosa), Fusarium basal rot (Fusarium oxysoporum, Fusariumsolani);Diseases of soybean: purple stain (Cercospora kikuchii), anthracnose(Elsinoe glycines), pod and stem blight (Diaporthe phaseolorum var.Sojae), brown spot (Septoria glycines), leaf spot (Cercospora sojina),rust (Phakopsora pachyrhizi), Fusarium blight (Phytophthora sojae),damping-off (Rhizoctonia solani) (Pythium spp.), root necrosis(Rhizoctonia solani), Fusarium root necrosis (Fusarium solani),anthracnose (Colletotrichum truncatum), Fusarium blight (Fusariumoxysporum, F. avenaceum, F. roseum), Sclerotinia rot (Sclerotiniasclerotiorum), gray mold (Botrytis cinerea);Diseases of adzuki bean: gray mold (Botrytis cinerea), Sclerotinia rot(Sclerotinia sclerotiorum), rust (Uromyces phaseoli), anthracnose(Coletotrichum phaseolorum);Diseases of kidney bean: gray mold (Botrytis cinerea), Sclerotinia rot(Sclerotinia sclerotiorum), anthracnose (Colletotrichum lindemthianum),Fusarium wilt (Fusarium oxysporum), rust (Uromyces phaseoli), angularleaf spot (Phaeoisariopsis griseola), Rhizoctonia root necrosis(Rhizoctonia solani), aphanomyces root necrosis (Aphanomyces euteiches);Diseases of peanut: leaf spot (Cercospora personata), brown leaf spot(Cercospora arachidicola), southern blight (Sclerotium rolfsii);Diseases of pea: gray mold (Botrytis cinerea), powdery mildew (Erysiphepisi), root necrosis (Fusarium solani f. Sp. Pisi);Diseases of potato: early blight (Alternaria solani), late blight(Phytophthora infestans), powdery scab (Spongospora subterranea), pinkrot (Phytophthora erythroseptica);Diseases of strawberry: gray mold (Botrytis cinerea), powdery mildew(Sphaerotheca hamuli), anthracnose (Glomerella cingulata);Diseases of tea: net blister blight (Exobasidium reticulatum), whitescab (Elsinoe leucospila), gray blight (Pestalotiopsis sp.), anthracnose(Colletotrichum theae-sinensis);Diseases of cotton: Fusarium wilt (Fusarium oxysporum), Fusarium wilt(Rhizoctonia solani);Diseases of tobacco: brown spot (Alternaria longipes), powdery mildew(Erysiphe cichoracearum), anthracnose (Colletotrichum tabacum), downymildew (Peronospora tabacina), black shank (Phytophthora nicotianae);Diseases of sugar beet: brown leaf spot (Cercospora beticola), leafblight (Thanatephorus cucumeris), root necrosis (Thanatephoruscucumeris), aphanomyces root rot (Aphanomyces cochlioides);Diseases of rose: scab (Diplocarpon rosae), powdery mildew (Sphaerothecapannosa), downy mildew (Peronospora sparsa), gray mold (Botrytiscinerea);Diseases of Chrysanthemum: brown leaf spot (Septoriachrysanthemi-indici), rust (Septoria chrysanthemi-indici), downy mildew(Bremia lactucae);Diseases of radish: alternaria leaf spot (Alternaria brassicicola);Disease of turfgrass: dollar spot (Sclerotinia homeocarpa), brown patchand large patch (Rhizoctonia solani);Diseases of banana: Sigatoka disease (Mycosphaerella fijiensis,Mycosphaerella musicola, Pseudocercospora musae);Diseases of sunflower: downy mildew (Plasmopara halstedii), alternarialeaf spot (Alternaria helianthi), southern blight (Sclerotium rolfsii),damping-off (Rhizoctonia solani), Sclerotinia rot (Sclerotiniasclerotiorum), rust (Puccinia helianthi);Diseases of various plants: diseases caused by Pythium spp. (Pythiumaphanidermatum, Pythium debarianum, Pythium graminicola, Pythiumirregulare, Pythium ultimum), gray mold (Botrytis cinerea), Sclerotiniarot (Sclerotinia sclerotiorum); damping-off (Rhizoctonia solani).

The present invention can be applied preferably to bacteria in the genusof Pythium spp., Rhizoctonia spp., and Fusarium spp., particularly,Pythium spp. and Rhizoctonia spp.

EXAMPLES

The invention is described in more detail with reference to thefollowing Preparation Examples, Formulation Examples, Seed TreatmentExamples, and Test Examples, which are not intended to limit the scopeof the present invention. The term “part” means “part by weight” unlessotherwise specified.

Preparation Example 1

A culture broth of the present bacterial strain, which has been culturedby a known technique, is centrifuged according to an ordinary method toseparate into a supernatant and a precipitate. The supernatant isremoved, and the precipitate is washed with sterilized water to obtain abacterial mass. The obtained bacterial mass is suspended in water, driedon spray drier, and the resultant dried product is pulverized to obtaina powder of the present bacterial strain.

Preparation Example 2

A culture broth of the present bacterial strain, which has been culturedby a known technique, is frozen at −80° C., freeze-dried and pulverizedto obtain a powder of the present bacterial strain.

Preparation Example 3

In a 500 mL Erlenmeyer flask with baffle, a platinum loop scraping ofthe present bacterial strain, which have been cultured in TSA (an agarmedium containing 15 g/L of casein peptone, 5 g/L of soybean peptone, 5g/L of sodium chloride, and 15 g/L of agar), are inoculated to a liquidmedium containing 200 mL TSB (a liquid medium containing 17 g/L ofcasein peptone, 3 g/L of soybean peptone, 2.5 g/L of glucose, 5 g/L ofsodium chloride and 2.5 g/L of K₂HPO₄) and incubated at 30° C. for 12hours to 24 hours to obtain a liquid culture. In a 500 mL volumeErlenmeyer flask with baffle, 2 mL of the liquid culture is inoculatedto 200 mL of a fresh TSB and cultured with shaking for 24 hours to 48hours to obtain a liquid culture of the present bacterial strain(hereinafter referred to as Liquid Culture a). The Liquid Culture a iscentrifuged according to a conventional manner to separate into asupernatant and precipitate. After removing the supernatant, theprecipitate is washed with sterile water and centrifuged. Thesupernatant is removed to obtain bacterial cells of the presentbacterial strain.

Preparation Example 4

The bacterial cells of the present bacterial strain obtained inPreparation Example 3 are suspended in water, dried on spray drier, andpulverized the resulting dried product to obtain a powder of the presentbacterial strain.

Preparation Example 5

The Liquid Culture a is obtained as described in Preparation 3. TheLiquid Culture a is frozen at −80° C., and freeze-dried and pulverizedto obtain a powder of the present bacterial strain.

Preparation Example 6

In a Erlenmeyer flask with baffle, a platinum loop scraping of thepresent bacterial strain, which have been cultured in TSA (an agarmedium containing 15 g/L of casein peptone, 5 g/L of soybean peptone, 5g/L of sodium chloride, and 15 g/L of agar), were inoculated to a liquidmedium containing 200 mL TSB (a liquid medium containing 17 g/L ofcasein peptone, 3 g/L of soybean peptone, 2.5 g/L of glucose, 5 g/L ofsodium chloride and 2.5 g/L of K₂HPO₄) and incubated at 30° C. for 23hours to obtain a liquid culture. The liquid culture (2% (v/v)) wasinoculated to a fresh TSB in a Erlenmeyer flask with baffle and culturedat 30° C. with shaking for 43 hours to obtain a liquid culture of thepresent bacterial strain (hereinafter referred to as Liquid Culture b).The liquid culture b was centrifuged at 1900×g for 10 min to separateinto a supernatant and a precipitate. After removing the supernatant,the precipitate was washed with sterilized water and centrifuged at1900×g for 10 min. The supernatant is removed to obtain 3.8×10¹¹ cfu/gof bacterial cells of the present bacterial strain.

Preparation Example 7

The bacterial cells of the present bacterial strain obtained asdescribed in Preparation Example 6 were frozen at −80° C. andfreeze-dried. The dried product obtained thus by freeze-drying waspulverized using scoopula to obtain 2.8×10¹² cfu/g of a powder of thepresent bacterial strain.

Formulation Example 1

To a mixture containing 3 parts of ethaboxam, 5 parts of white carbon, 8parts of sodium lignin sulfonate and 2 parts of sodium alkyl naphthalenesulfonate are added the powder of the present bacterial strain obtainedas described in Preparation Example 1 or 2, in an amount of 1×10¹⁰ cfuper 1 g of the formulation, and diatomaceous earth to 100 parts,followed by mixing and grinding to obtain wettable powder.

Formulation Example 2

To a mixture containing 1 part of tolclofos-methyl, 5 parts of whitecarbon, 8 parts of sodium lignin sulfonate, 2 parts of sodium alkylnaphthalene sulfonate are added the powder of the present bacterialstrain obtained as described in Preparation Example 1 or 2, in an amountof 1×10¹⁰ cfu per 1 g of the formulation, and diatomaceous earth to 100parts, followed by mixing and grinding to obtain wettable powder.

Formulation Example 3

To a mixture containing 3 parts of metalaxyl, 5 parts of white carbon, 8parts of sodium lignin sulfonate, 2 parts of sodium alkyl naphthalenesulfonate are added the powder of the present bacterial strain obtainedas described in Preparation Example 1 or 2, in an amount of 1×10¹⁰ cfuper 1 g of the formulation, and diatomaceous earth to 100 parts,followed by mixing and grinding to obtain wettable powder.

Formulation Example 4

To a mixture containing 1 part of fludioxonil, 5 parts of white carbon,8 parts of sodium lignin sulfonate, 2 parts of sodium alkyl naphthalenesulfonate are added the powder of the present bacterial strain obtainedas described in Preparation Example 1 or 2, in an amount of 1×10¹⁰ cfuper 1 g of the formulation, and diatomaceous earth to 100 parts,followed by mixing and grinding to obtain wettable powder.

Formulation Example 5

To a mixture containing 1 part of the present carboxamide compound (1),5 parts of white carbon, 8 parts of sodium lignin sulfonate, 2 parts ofsodium alkyl naphthalene sulfonate are added the powder of the presentbacterial strain obtained as described in Preparation Example 1 or 2, inan amount of 1×10¹⁰ cfu per 1 g of the formulation, and diatomaceousearth to 100 parts, followed by mixing and grinding to obtain wettablepowder.

Formulation Example 6

To a mixture containing 15 parts of ethaboxam and 30 parts of whitecarbon containing 30% by weight of polyoxyethylene alkyl ether sulfateammonium salt are added the powder of the present bacterial strainobtained as described in Preparation Example 1 or 2, in an amount of1×10¹⁰ cfu per 1 g of the formulation, and water to 100 parts, followedby wet-milling to finely milled to obtain a flowable formulation.

Formulation Example 7

To a mixture containing 5 parts of tolclofos-methyl and 30 parts ofwhite carbon containing 30% by weight of polyoxyethylene alkyl ethersulfate ammonium salt are added the powder of the present bacterialstrain obtained as described in Preparation Example 1 or 2, in an amountof 1×10¹⁰ cfu per 1 g of the formulation, and water to 100 parts,followed by wet-milling to finely milled to obtain a flowableformulation.

Formulation Example 8

To a mixture containing 15 parts of metalaxyl and 30 parts of whitecarbon containing 30% by weight of polyoxyethylene alkyl ether sulfateammonium salt are added the powder of the present bacterial strainobtained as described in Preparation Example 1 or 2, in an amount of1×10¹⁰ cfu per 1 g of the formulation, and water to 100 parts, followedby wet-milling to finely milled to obtain a flowable formulation.

Formulation Example 9

To a mixture containing 5 parts of fludioxonil and 30 parts of whitecarbon containing 30% by weight of polyoxyethylene alkyl ether sulfateammonium salt are added the powder of the present bacterial strainobtained as described in Preparation Example 1 or 2, in an amount of1×10¹⁰ cfu per 1 g of the formulation, and water to 100 parts, followedby wet-milling to finely milled to obtain a flowable formulation.

Formulation Example 10

Mixing 32.8 parts of water, 37 parts of the carboxamide compound (1),4.6 parts of polyoxyethylene tristyryl phenyl ether phosphoric acidester potassium salt/propylene glycol (40/60) and 0.2 part of anemulsion-type silicone antifoamer, followed by finely milling bywet-milling to obtain Suspension A. To 19 parts of water are added 0.4part of magnesium aluminum silicate, 4.6 parts of propylene glycol, 0.2part of xanthan gum, and 0.3 part of a preservative containing1,2-benzisothiazolin-3-one as an active ingredient, followed by stirringto obtain Thickening Agent Solution A. The Suspension A and theThickening Agent Solution A are mixed to obtain a formulation. Thepowder of the present bacterial strain obtained as described inPreparation Example 1 or 2, in an amount of 4×10¹¹ cfu per 1 g of theformulation, and water to 100 parts are added and finely milled bywet-milling to obtain a flowable formulation.

Formulation Example 11

To a mixture containing 5 parts of white carbon, 8 parts of sodiumlignin sulfonate, and 2 parts of sodium alkyl naphthalene sulfonate areadded the bacterial cells or the powder of the present bacterial strainobtained as described in any one of Preparation Examples 3 to 5, in anamount of 1×10¹⁰ cfu per 1 g of the formulation, and diatomaceous earthto 100 parts to obtain a mixture. The mixture is milled to obtainwettable powder of the present bacterial strain.

Formulation Example 12

To 30 parts of white carbon containing 30% by weight of polyoxyethylenealkyl ether sulfate ammonium salt are added the bacterial cells orpowder of the present bacterial strain obtained as described in any oneof Preparation Examples 3 to 5, in an amount of 1×10¹⁰ cfu or 1×10¹² cfuper 1 g of the formulation, and water to 100 parts to obtain a mixture.The mixture is finely milled by wet-milling to obtain a flowableformulation of the present bacterial strain.

Formulation Example 13

To 30 parts of white carbon containing 30% by weight of polyoxyethylenealkyl ether sulfate ammonium salt were added the powder of the presentbacterial strain obtained in Preparation Example 7, in an amount of1×10¹⁰ cfu or 1×10¹² cfu per 1 g of the formulation, and water to 100parts to obtain a mixture. The mixture was finely milled by wet-millingto obtain a flowable formulation of the present bacterial strain.

Seed Treatment Examples are provided below.

Seed Treatment Example 1

To a mixture containing 5 parts of white carbon, 8 parts of sodiumlignin sulfonate and 2 parts of sodium alkyl naphthalene sulfonate areadded the powder of the present bacterial strain obtained as describedin Preparation Example 1, in an amount of 1×10¹⁰ cfu per 1 g of theobtained formulation, and diatomaceous earth to 100 parts, followed bymixing and grinding to obtain wettable powder of the present bacterialstrain.

Soybean seeds are treated by smearing treatment with tolclofos-methylflowable formulation (42% flowable formulation, trade name: Rizolex,Valent U.S.A. Corporation) in an amount of 0.1 g of tolclofos-methyl per1 kg of the soybean seeds. The soybean seeds thus treated withtolclofos-methyl are treated by wet powder coating with the wettable ofthe present bacterial strain in an amount of 1×10¹⁰ cfu of the presentbacterial strain per 1 kg of said soybean seeds.

Seed Treatment Example 2

Soybean seeds are treated by smearing treatment with metalaxyl flowableformulation (33.92% flowable formulation, trade name: Apron XL, SyngentaCrop Protection LLC) in an amount of 0.3 g of metalaxyl per 1 kg of thesoybean seeds. The soybean seeds thus treated with metalaxyl are treatedby wet powder coating with wettable powder of the present bacterialstrain prepared as described in Seed Treatment Example 1, in an amountof 1×10¹⁰ cfu/kg of the present bacterial strain per 1 kg of saidsoybean seeds.

Seed Treatment Example 3

To 30 parts of white carbon containing 30 parts of polyoxyethylene alkylether sulfate ammonium salt are added a powder of the present bacterialstrain obtained as described in Preparation Example 2, in an amount of1×10¹⁰ cfu per 1 g of the formulation, and water to 100 parts, and themixture is finely milled by wet-milling to obtain a flowable formulationof the present bacterial strain.

Soybean seeds are treated by smearing treatment with a liquid mixturecontaining the flowable formulation of the present bacterial strain(1×10¹⁰ cfu per 1 kg of the soybean seeds) and ethaboxam flowableformulation (38.3% flowable formulation, trade name: Intego Solo, ValentU.S.A. Corporation), in an amount of 0.3 g of ethoboxam per 1 kg of thesoybean seeds.

Seed Treatment Example 4

Soybean seeds are treated by smearing treatment with a solution of thecarboxamide compound (1) in acetone/Tween 20 (weight ratio=95:5) dilutedwith water, in an amount of 0.1 g of the carboxamide compound (1) per 1kg of the soybean seeds.

The soybean seeds thus treated with the carboxamide compound (1) aretreated by smearing treatment with a flowable formulation of the presentbacterial strain prepared as described in Seed Treatment Example 3, inan amount of 1×10¹⁰ cfu of the present bacterial strain per 1 kg of saidsoybean seeds.

Seed Treatment Example 5

Corn seeds are treated by smearing treatment with the flowableformulation of the present bacterial strain and metalaxyl prepared asdescribed in Formulation Example 8, in an amount of 2×10¹⁰ cfu of thepresent bacterial strain and 0.3 g of metalaxyl per 1 kg of the cornseeds.

Seed Treatment Example 6

Soybean seeds are treated by smearing treatment with the flowableformulation of the present bacterial strain and fludioxonil prepared asdescribed in Formulation Example 9, in an amount of 2×10¹⁰ cfu of thepresent bacterial strain and 0.1 g of fludioxonil per 1 kg of thesoybean seeds.

Seed Treatment Example 7

Soybean seeds are treated by smearing treatment with tolclofos-methylflowable formulation (42% flowable formulation, trade name: Rizolex,Valent U.S.A. Corporation), in an amount of 0.1 g of tolclofos-methylper 1 kg of the soybean seeds. The soybean seeds thus treated withtolclofos-methyl are treated by wet powder coating with wettable powderof the present bacterial strain obtained as described in FormulationExample 11, in an amount of 1×10¹⁰ cfu of the present bacterial strainper 1 kg of said soybean seeds.

Seed Treatment Example 8

Soybean seeds are treated by smearing treatment with a liquid mixturecontaining a flowable formulation of the present bacterial strainobtained as described in Formulation Example 12, in an amount of 1×10¹⁰cfu of the present bacterial strain per 1 kg of the soybean seeds, andethaboxam flowable formulation (38.3% flowable formulation, trade name:Intego Solo, Valent U.S.A. Corporation) in an amount of 0.3 g ofethoboxam for 1 kg of the soybean seeds.

Test Examples are provided below.

Test Example 1

In a rotary seed treatment machine (trade name: HEGE11, manufactured byWINTERSTEIGER), corn seeds (variety: yellow dent corn) are treated bysmearing treatment with a liquid mixture containing a flowableformulation of the present bacterial strain as obtained in the SeedTreatment Example 3 (adjusted to 1×10¹⁰ cfu per 1 kg of the corn seeds)and ethaboxam flowable formulation (38.3% flowable formulation, tradename: Intego Solo, Valent U.S.A. Corporation, adjusted to 0.3 g ofethoboxam per 1 kg of the soybean seeds).

A plastic pot is filled with a soil, and then, the treated seeds areseeded and covered with a soil, which has been mixed with damping-offfungus (Pythium spp.) cultured in a bran medium. Cultivation is carriedout in a greenhouse under irrigation (“treated compartment”). The plantsare investigated 20 days after for the number of diseased plants, andthe disease incidence is calculated by the following “Equation 1”.Separately, untreated corn seeds are seeded, covered with a soil andcultivated in a similar manner as described above for “treatedcompartment” (“untreated compartment”). The plants are investigated 20days after for the number of diseased plants, and the disease incidenceis calculated by the following “Equation 1”. Based on the diseaseincidences of the treated compartment and the untreated compartment, thecontrol value of the treated compartment is calculated by the followingequation “Equation 2”, and the treated compartment is confirmed to havea good control effect on plant disease.Disease incidence (%)=100×(number of diseased plant/total number ofseeded seeds):  Equation 1Control value (%)=100×[(disease incidence in untreatedcompartment−disease incidence in treated compartment)/disease incidencein untreated compartment]:  Equation 2

The composition of the invention shows a significantly highercontrolling effect.

Test Example 2

In a rotary seed treatment machine (trade name: HEGE11, manufactured byWINTERSTEIGER), soybean seeds (variety:“Hatayutaka”) are treated bysmearing treatment with a liquid mixture containing a flowableformulation of the present bacterial strain as obtained in the SeedTreatment Example 3 (adjusted to 1×10¹⁰ cfu per 1 kg of the corn seeds)and metalaxyl flowable formulation (33.92% flowable formulation, tradename: Apron XL, Syngenta Crop Protection LLC, adjusted to 0.3 g ofmetalaxyl per 1 kg of the soybean seeds).

A plastic pot is filled with a soil, and then, the treated seeds areseeded and covered with a soil, which has been mixed with damping-offfungus (Pythium spp.) cultured in a bran medium. Cultivation is carriedout in a greenhouse under irrigation (“treated compartment”). The plantsare investigated 20 days after for the number of diseased plants, andthe disease incidence is calculated by the following “Equation 1”.Separately, untreated soybean seeds are seeded, covered with a soil andcultivated in a similar manner as described above for “treatedcompartment” (“untreated compartment”). The plants are investigated 20days after for the number of diseased plants, and the disease incidenceis calculated by the following “Equation 1”. Based on the diseaseincidences of the treated compartment and the untreated compartment, thecontrol value of the treated compartment is calculated by the followingequation “Equation 2”, and the treated compartment is confirmed to havea good control effect on plant disease.Disease incidence (%)=100×(number of diseased plant/total number ofseeded seeds):  Equation 1Control value (%)=100×[(disease incidence in untreated compartmentdisease incidence in treated compartment)/disease incidence in untreatedcompartment]:  Equation 2

The composition of the invention shows a significantly highercontrolling effect.

Test Example 3

Soybean seeds (variety:“Hatayutaka”) are treated by wet powder coatingtreatment with the wettable powder of the present bacterial strain andtolclofos-methyl prepared in Formulation Example 2 (adjusted to 1×10¹¹cfu of the present bacterial strain and 0.1 g of tolclofos-methyl per 1kg of the soybean seeds), with the wettable powder of the presentbacterial strain and fludioxonil prepared in Formulation Example 4(adjusted to 1×10¹¹ cfu of the present bacterial strain and 0.1 g offludioxonil per 1 kg of the soybean seeds) or with the wettable powderof the present bacterial strain and the carboxamide compound (1)prepared in Formulation Example 5 (adjusted to 1×10¹¹ cfu of the presentbacterial strain and 0.1 g of the carboxamide compound (1) per 1 kg ofthe soybean seeds).

A plastic pot is filled with a soil, and then, the treated seeds areseeded and covered with a soil, which has been mixed with damping-offfungus (Rhizoctonia solani) cultured in a bran medium. Cultivation iscarried out in a greenhouse under irrigation (“treated compartment”).The plants are investigated 20 days after for the number of diseasedplants, and the disease incidence is calculated by the following“Equation 1”. Separately, untreated soybean seeds are seeded, coveredwith a soil and cultivated in a similar manner as described above for“treated compartment” (“untreated compartment”). The plants areinvestigated 20 days after for the number of diseased plants, and thedisease incidence is calculated by the following “Equation 1”. Based onthe disease incidences of the treated compartment and the untreatedcompartment, the control value of the treated compartment is calculatedby the following equation “Equation 2”, and the treated compartment isconfirmed to have a good control effect on plant disease.Disease incidence (%)=100×(number of diseased plant/total number ofseeded seeds):  Equation 1Control value (%)=100×[(disease incidence in untreated compartmentdisease incidence in treated compartment)/disease incidence in untreatedcompartment]:  Equation 2

The composition of the invention shows a significantly highercontrolling effect.

Test Example 4

Soybean seeds (variety:“Hatayutaka”) are treated by wet powder coatingtreatment with the wettable powder of the present bacterial strain andethaboxam prepared in Formulation Example 1 (adjusted to 1×10¹¹ cfu ofthe present bacterial strain and 0.3 g of ethaboxam per 1 kg of thesoybean seeds) or with the wettable powder of the present bacterialstrain and metalaxyl prepared in Formulation Example 3 (adjusted to1×10¹¹ cfu of the present bacterial strain and 0.3 g of metalaxyl per 1kg of the soybean seeds).

A plastic pot is filled with a soil, and then, the treated seeds areseeded and covered with a soil, which has been mixed with damping-offfungus (Pythium spp.) cultured in a bran medium. Cultivation is carriedout in a greenhouse under irrigation (“treated compartment”). The plantsare investigated 20 days after for the number of diseased plants, andthe disease incidence is calculated by the following “Equation 1”.Separately, untreated soybean seeds are seeded, covered with a soil andcultivated in a similar manner as described above for “treatedcompartment” (“untreated compartment”). The plants are investigated 20days after for the number of diseased plants, and the disease incidenceis calculated by the following “Equation 1”. Based on the diseaseincidences of the treated compartment and the untreated compartment, thecontrol value of the treated compartment is calculated by the followingequation “Equation 2”, and the treated compartment is confirmed to havea good control effect on plant disease.Disease incidence (%)=100×(number of diseased plant/total number ofseeded seeds):  Equation 1Control value (%)=100×[(disease incidence in untreated compartmentdisease incidence in treated compartment)/disease incidence in untreatedcompartment]:  Equation 2

The composition of the invention shows a significantly highercontrolling effect.

Test Example 5

In a rotary seed treatment machine (trade name: HEGE11, manufactured byWINTERSTEIGER), soybean seeds are treated by smearing treatment with aflowable formulation of the present bacterial strain andtolclofos-methyl prepared in Formulation Example 7 (adjusted to 2×10¹⁰cfu of the present bacterial strain and 0.1 g of tolclofos-methyl per 1kg of the soybean seeds), with a flowable formulation of the presentbacterial strain and fludioxonil prepared in Formulation Example 9(adjusted to 2×10¹⁰ cfu of the present bacterial strain and 0.3 g offludioxonil per 1 kg of the soybean seeds), or with a flowableformulation of the present bacterial strain and the carboxamide compound(1) prepared in Formulation Example 10 (adjusted to 2×10¹⁰ cfu of thepresent bacterial strain and 0.1 g of the carboxamide compound (1) per 1kg of the soybean seeds).

A plastic pot is filled with a soil, and then, the treated seeds areseeded and covered with a soil, which has been mixed with damping-offfungus (Rhizoctonia solani) cultured in a bran medium. Cultivation iscarried out in a greenhouse under irrigation (“treated compartment”).The plants are investigated 20 days after for the number of diseasedplants, and the disease incidence is calculated by the following“Equation 1”. Separately, untreated soybean seeds are seeded, coveredwith a soil and cultivated in a similar manner as described above for“treated compartment” (“untreated compartment”). The plants areinvestigated 20 days after for the number of diseased plants, and thedisease incidence is calculated by the following “Equation 1”. Based onthe disease incidences of the treated compartment and the untreatedcompartment, the control value of the treated compartment is calculatedby the following equation “Equation 2”, and the treated compartment isconfirmed to have a good control effect on plant disease.Disease incidence (%)=100×(number of diseased plant/total number ofseeded seeds):  Equation 1Control value (%)=100×[(disease incidence in untreated compartmentdisease incidence in treated compartment)/disease incidence in untreatedcompartment]:  Equation 2

The composition of the invention shows a significantly highercontrolling effect.

Test Example 6

In a rotary seed treatment machine (trade name: HEGE11, manufactured byWINTERSTEIGER), soybean seeds are treated by smearing treatment with theflowable formulation of the present bacterial strain and ethaboxamprepared in Formulation Example 6 (adjusted to 2×10¹⁰ cfu of the presentbacterial strain and 0.3 g of ethaboxam per 1 kg of the soybean seeds)or with the flowable formulation of the present bacterial strain andmetalaxyl prepared in Formulation Example 8 (adjusted to 2×10¹⁰ cfu ofthe present bacterial strain and 0.3 g of metalaxyl per 1 kg of thesoybean seeds).

A plastic pot is filled with a soil, and then, the treated seeds areseeded and covered with a soil, which has been mixed with damping-offfungus (Pythium spp.) cultured in a bran medium. Cultivation is carriedout in a greenhouse under irrigation (“treated compartment”). The plantsare investigated 20 days after for the number of diseased plants, andthe disease incidence is calculated by the following “Equation 1”.Separately, untreated soybean seeds are seeded, covered with a soil andcultivated in a similar manner as described above for “treatedcompartment” (“untreated compartment”). The plants are investigated 20days after for the number of diseased plants, and the disease incidenceis calculated by the following “Equation 1”. Based on the diseaseincidences of the treated compartment and the untreated compartment, thecontrol value of the treated compartment is calculated by the followingequation “Equation 2”, and the treated compartment is confirmed to havea good control effect on plant disease.Disease incidence (%)=100×(number of diseased plant/total number ofseeded seeds):  Equation 1Control value (%)=100×[(disease incidence in untreated compartmentdisease incidence in treated compartment)/disease incidence in untreatedcompartment]:  Equation 2

The composition of the invention shows a significantly highercontrolling effect.

Test Example 7

In a rotary seed treatment machine (trade name: HEGE11, manufactured byWINTERSTEIGER), soybean seeds are treated by smearing treatmentrespectively with a chemical liquid, which has been prepared bydissolving a tolclofos-methyl flowable formulation (42% flowableformulation, trade name: Rizolex, Valent U.S.A. Corporation) in anamount of 0.1 g of tolclofos-methyl per 1 kg of the soybean seeds or afludioxonil flowable formulation (10% flowable formulation, trade name:Maxim100, Syngenta Crop Protection LLC) in an amount of 0.1 g offludioxonil per 1 kg of the soybean seeds, or the carboxamide compound(1) in an amount of 0.1 g of fludioxonil per 1 kg of the soybean seeds,in acetone/Tween 20 (weight ratio=95:5), followed by diluting withwater. In a rotary seed treatment machine (trade name: HEGE11,manufactured by WINTERSTEIGER), the soybean seeds thus treated withethaboxam, tolclofos-methyl, fludioxonil or the carboxamide compound(1), respectively, are treated by wet powder coating with the wettablepowder of the present bacterial strain obtained in Seed TreatmentExample 1 in an amount of 1×10¹⁰ cfu of the present bacterial strain per1 kg of said soybean seeds.

A plastic pot is filled with a soil, and then, the treated seeds areseeded and covered with a soil, which has been mixed with damping-offfungus (Rhizoctonia solani) cultured in a bran medium. Cultivation iscarried out in a greenhouse under irrigation (“treated compartment”).The plants are investigated 20 days after for the number of diseasedplants, and the disease incidence is calculated by the following“Equation 1”. Separately, untreated soybean seeds are seeded, coveredwith a soil and cultivated in a similar manner as described above for“treated compartment” (“untreated compartment”). The plants areinvestigated 20 days after for the number of diseased plants, and thedisease incidence is calculated by the following “Equation 1”. Based onthe disease incidences of the treated compartment and the untreatedcompartment, the control value of the treated compartment is calculatedby the following equation “Equation 2”, and the treated compartment isconfirmed to have a good control effect on plant disease.Disease incidence (%)=100×(number of diseased plant/total number ofseeded seeds):  Equation 1Control value (%)=100×[(disease incidence in untreated compartmentdisease incidence in treated compartment)/disease incidence in untreatedcompartment]:  Equation 2

The composition of the invention shows a significantly highercontrolling effect.

Test Example 8

In a rotary seed treatment machine (trade name: HEGE11, manufactured byWINTERSTEIGER), soybean seeds are treated by smearing treatment with asolution prepared by dissolving a metalaxyl flowable formulation (33.92%flowable formulation, trade name: Apron XL, Syngenta Crop ProtectionLLC, adjusted to 0.3 g of metalaxyl per 1 kg of the soybean seeds). In arotary seed treatment machine (trade name: HEGE11, manufactured byWINTERSTEIGER), the soybean seeds thus treated with metalaxyl aretreated by wet powder coating with the wettable powder of the presentbacterial strain obtained in Seed Treatment Example 1 in an amount of1×10¹⁰ cfu of the present bacterial strain per 1 kg of said soybeanseeds.

A plastic pot is filled with a soil, and then, the treated seeds areseeded and covered with a soil, which has been mixed with damping-offfungus (Pythium spp.) cultured in a bran medium. Cultivation is carriedout in a greenhouse under irrigation (“treated compartment”). The plantsare investigated 20 days after for the number of diseased plants, andthe disease incidence is calculated by the following “Equation 1”.Separately, untreated soybean seeds are seeded, covered with a soil andcultivated in a similar manner as described above for “treatedcompartment” (“untreated compartment”). The plants are investigated 20days after for the number of diseased plants, and the disease incidenceis calculated by the following “Equation 1”. Based on the diseaseincidences of the treated compartment and the untreated compartment, thecontrol value of the treated compartment is calculated by the followingequation “Equation 2”, and the treated compartment is confirmed to havea good control effect on plant disease.Disease incidence (%)=100×(number of diseased plant/total number ofseeded seeds):  Equation 1Control value (%)=100×[(disease incidence in untreated compartmentdisease incidence in treated compartment)/disease incidence in untreatedcompartment]:  Equation 2

The composition of the invention shows a significantly highercontrolling effect.

Test Example 9

In a rotary seed treatment machine (trade name: HEGE11, manufactured byWINTERSTEIGER), soybean seeds are treated by smearing treatment with theflowable formulation of the present bacterial strain obtained in SeedTreatment Example 3 (adjusted to 1×10¹⁰ cfu of the present bacterialstrain per 1 kg of the soybean seeds). In a rotary seed treatmentmachine (trade name: HEGE11, manufactured by WINTERSTEIGER), soybeanseeds thus treated by smearing treatment are treated with a chemicalliquid, which has been prepared by dissolving a tolclofos-methylflowable formulation (42% flowable formulation, trade name: Rizolex,Valent U.S.A. Corporation, adjusted to 0.1 g of tolclofos-methyl per 1kg of the soybean seeds) or a fludioxonil flowable formulation (10%flowable formulation, trade name: Maxim100, Syngenta Crop ProtectionLLC, adjusted to 0.1 g of fludioxonil per 1 kg of the soybean seeds) orthe carboxamide compound (1) (adjusted to 0.1 g of fludioxonil per 1 kgof the soybean seeds) in acetone/Tween 20 (weight ratio=95:5), followedby diluting with water, respectively.

A plastic pot is filled with a soil, and then, the treated seeds areseeded and covered with a soil, which has been mixed with damping-offfungus (Rhizoctonia solani) cultured in a bran medium. Cultivation iscarried out in a greenhouse under irrigation (“treated compartment”).The plants are investigated 20 days after for the number of diseasedplants, and the disease incidence is calculated by the following“Equation 1”. Separately, untreated soybean seeds are seeded, coveredwith a soil and cultivated in a similar manner as described above for“treated compartment” (“untreated compartment”). The plants areinvestigated 20 days after for the number of diseased plants, and thedisease incidence is calculated by the following “Equation 1”. Based onthe disease incidences of the treated compartment and the untreatedcompartment, the control value of the treated compartment is calculatedby the following equation “Equation 2”, and the treated compartment isconfirmed to have a good control effect on plant disease.Disease incidence (%)=100×(number of diseased plant/total number ofseeded seeds):  Equation 1Control value (%)=100×[(disease incidence in untreated compartmentdisease incidence in treated compartment)/disease incidence in untreatedcompartment]:  Equation 2

The composition of the invention shows a significantly highercontrolling effect.

Test Example 10

In a rotary seed treatment machine (trade name: HEGE11, manufactured byWINTERSTEIGER), soybean seeds are treated by smearing treatment with aflowable formulation of the present bacterial strain obtained in SeedTreatment Example 3 (adjusted to 1×10¹⁰ cfu of the present bacterialstrain per 1 kg of the soybean seeds). In a rotary seed treatmentmachine (trade name: HEGE11, manufactured by WINTERSTEIGER), soybeanseeds thus treated by smearing treatment are treated with a chemicalliquid, which has been prepared by dissolving an ethaboxam flowableformulation (38.3% flowable formulation, trade name: Intego Solo, ValentU.S.A. Corporation, adjusted to 0.3 g of ethaboxam per 1 kg of thesoybean seeds) or a metalaxyl flowable formulation (33.92% flowableformulation, trade name: Apron XL, using the Syngenta Crop ProtectionLLC, adjusted to 0.3 g of metalaxyl per 1 kg of the soybean seeds),respectively.

A plastic pot is filled with a soil, and then, the treated seeds areseeded and covered with a soil, which has been mixed with damping-offfungus (Pythium spp.) cultured in a bran medium. Cultivation is carriedout in a greenhouse under irrigation (“treated compartment”). The plantsare investigated 20 days after for the number of diseased plants, andthe disease incidence is calculated by the following “Equation 1”.Separately, untreated soybean seeds are seeded, covered with a soil andcultivated in a similar manner as described above for “treatedcompartment” (“untreated compartment”). The plants are investigated 20days after for the number of diseased plants, and the disease incidenceis calculated by the following “Equation 1”. Based on the diseaseincidences of the treated compartment and the untreated compartment, thecontrol value of the treated compartment is calculated by the followingequation “Equation 2”, and the treated compartment is confirmed to havea good control effect on plant disease.Disease incidence (%)=100×(number of diseased plant/total number ofseeded seeds):  Equation 1Control value (%)=100×[(disease incidence in untreated compartmentdisease incidence in treated compartment)/disease incidence in untreatedcompartment]:  Equation 2

The composition of the invention shows a significantly highercontrolling effect.

Test Example 11

A chemical liquid is prepared by diluting ethaboxam flowable formulation(12.5% flowable formulation, trade name: Ethofin flowable, Nippon SodaCo., Ltd.) with water to 400 ppm of ethaboxam or dissolving metalaxyl inacetone/Tween 20 (weight ratio=95:5) and diluting with water to 400 ppmof metalaxyl, and followed by combining the solution with the equalvolume of a solution of the flowable formulation obtained in SeedTreatment Example 1 (adjusted to 2×10⁸ cfu of the present bacterialstrain).

The chemical liquid is sprayed to tomato plants (variety: Campari,4-leaf stage) in a sufficient amount. After air drying, the plants areinoculated with late blight fungus (Phytophthora infestans) and left tostand under moisture condition for 10 days.

The effect on the treated compartment is determined by the followingequation, based on the onset area rates of the treated compartment andthe untreated compartment.controlling effect=100×[1−(onset area rate of treatedcompartment)/(onset area rate of untreated compartment)]  Equation:

The composition of the invention shows a significantly highercontrolling effect.

Test Example 12

A chemical liquid is prepared by diluting fludioxonil flowableformulation (20% flowable formulation, trade name: Savior flowable,Syngenta Japan) with water to 400 ppm of fludioxonil or dissolving thecarboxamide compound (1) in acetone/Tween 20 (weight ratio=95:5) anddiluting with water to 400 ppm of the carboxamide compound (1), andfollowed by combining the solution with the equal volume of a solutionof the flowable formulation obtained in Seed Treatment Example 1(adjusted to 2×10⁸ cfu of the present bacterial strain).

The chemical liquid is sprayed to cucumber plants (variety: “SagamiHanjiro”, 3-leaf stage) in a sufficient amount. After air drying, theplants are inoculated with gray mold fungus (Botrytis cinerea) and leftto stand under moisture condition for 10 days.

The effect on the treated compartment is determined by the followingequation, based on the onset area rates of the treated compartment andthe untreated compartment.controlling effect=100×[1−(onset area rate of treatedcompartment)/(onset area rate of untreated compartment)]  Equation:

The composition of the invention shows a significantly highercontrolling effect.

Test Example 13

A chemical liquid is prepared by diluting tolclofos-methyl wettablepowder (50% wettable powder, trade name: Rizolex 50, Sumitomo ChemicalCo., Ltd.) with water to 400 ppm of tolclofos-methyl or dissolving thecarboxamide compound (1) in acetone/Tween 20 (weight ratio=95:5) anddiluting with water to 400 ppm of the carboxamide compound (1), andfollowed by combining the solution with the equal volume of a solutionof the flowable formulation obtained in Seed Treatment Example 1(adjusted to 2×10⁸ cfu of the present bacterial strain).

The chemical liquid is sprayed to sugar beet plants (variety:“Yukimaru”, growth stage) in a sufficient amount. After air drying, theplants are inoculated with leaf blight fungus (Thanatephorus cucumeris)and left to stand under moisture condition for 10 days.

The effect on the treated compartment is determined by the followingequation, based on the onset area rates of the treated compartment andthe untreated compartment.controlling effect=100×[1−(onset area rate of treatedcompartment)/(onset area rate of untreated compartment)]  Equation:

The composition of the invention shows a significantly highercontrolling effect.

Test Example 14

A chemical liquid is prepared by dissolving the carboxamide compound (1)in acetone/Tween 20 (weight ratio=95:5) and diluting with water to 400ppm of the carboxamide compound (1), and followed by combining thesolution with the equal volume of a solution of the flowable formulationobtained in Seed Treatment Example 1 (adjusted to 2×10⁸ cfu of thepresent bacterial strain).

The chemical liquid is sprayed to wheat plants (cultivar: Apogee, growthstage) in a sufficient amount. After air drying, the plants areinoculated with leaf blight fungus (Septoria tritici) and left to standunder moisture condition for 10 days.

The effect on the treated compartment is determined by the followingequation, based on the onset area rates of the treated compartment andthe untreated compartment.controlling effect=100×[1−(onset area rate of treatedcompartment)/(onset area rate of untreated compartment)]  Equation:

The composition of the invention shows a significantly highercontrolling effect.

Test Example 15

In a rotary seed treatment machine (trade name: HEGE11, manufactured byWINTERSTEIGER), corn seeds (variety: yellow dent corn) are treated bysmearing treatment using a flowable formulation of the present bacterialstrain as obtained in Formulation Example 12 and ethaboxam flowableformulation (38.3% flowable formulation, trade name: Intego Solo, ValentU.S.A. Corporation), so that the soybean seeds retain the presentbacterial strain and/or the compound in the amount shown in Table 1.

A plastic pot is filled with a soil, and then, the coon seeds, whichhave been treated with the present bacterial strain, compound or thepresent bacterial strain+compound as shown in Table 1, are seeded andcovered with a soil, which has been mixed with damping-off fungus(Pythium spp.) cultured in a bran medium or grass medium (“treatedcompartment”). Cultivation is carried out in a greenhouse underirrigation. The plants are investigated 20 days after for the number ofdiseased plants, and the disease incidence is calculated by thefollowing “Equation 1”. Similar procedures are conducted using untreatedcorn seeds, instead of the treated corn seeds, as described above forthe treated compartment (“untreated compartment”). The plants areinvestigated 20 days after for the number of diseased plants, and thedisease incidence is calculated by the following “Equation 1”. Based onthe disease incidences of the treated compartment and the untreatedcompartment, the control value of the treated compartment is calculatedby the following equation “Equation 2”, and the treated compartment isconfirmed to have a good control effect on plant disease.Disease incidence (%)=100×(number of diseased plant/total number ofseeded seeds):  Equation 1Control value (%)=100×[(disease incidence in untreated compartmentdisease incidence in treated compartment)/disease incidence in untreatedcompartment]:  Equation 2

The compartment treated with the composition of the invention shows asynergistic controlling effect, for each combination of the presentbacterial strain and the compound, compared with that of thecorresponding compartment treated solely with the present bacterialstrain or the compound.

TABLE 1 Retaining Bacteria/Compound Amount retained by Seeds (per Kg ofseeds) The Present Bacterial strain 1 × 10¹⁰ cfu The Present Bacterialstrain 1 × 10¹² cfu The Present Bacterial strain 1 × 10⁷ cfu Ethaboxam0.00625 g Ethaboxam 0.025 g Ethaboxam 0.1 g The Present Bacterial strain1 × 10¹⁰ cfu Ethaboxam 0.00625 g The Present Bacterial strain 1 × 10¹⁰cfu Ethaboxam 0.025 g The Present Bacterial strain 1 × 10¹⁰ cfuEthaboxam 0.1 g The Present Bacterial strain 1 × 10¹² cfu Ethaboxam0.00625 g The Present Bacterial strain 1 × 10¹² cfu Ethaboxam 0.025 gThe Present Bacterial strain 1 × 10⁷ cfu Ethaboxam 0.1 g

Test Example 16

Soybean seeds (variety:“Hatayutaka”) are treated by smearing treatmentusing wettable powder of the present bacterial strain as obtained inFormulation 11, a flowable formulation of the present bacterial strainas obtained in Formulation Example 12, ethaboxam flowable formulation(38.3% flowable formulation, trade name: Intego Solo, Valent U.S.A.Corporation) and metalaxyl flowable formulation (33.92% flowableformulation, trade name: Apron XL, Syngenta Crop Protection LLC), sothat the soybean seeds retain the present bacterial strain and/or thecompound in the amount shown in Table 2.

A plastic pot is filled with a soil, and then, the coon seeds, whichhave been treated with the present bacterial strain, compound or thepresent bacterial strain+compound as shown in Table 2, are seeded andcovered with a soil, which has been mixed with damping-off fungus(Pythium spp.) cultured in a bran medium or grass medium (“treatedcompartment”). Cultivation is carried out in a greenhouse underirrigation. The plants are investigated 20 days after for the number ofdiseased plants, and the disease incidence is calculated by thefollowing “Equation 1”. Similar procedures are conducted using untreatedcorn seeds, instead of the treated corn seeds, as described above forthe treated compartment (“untreated compartment”). The plants areinvestigated 20 days after for the number of diseased plants, and thedisease incidence is calculated by the following “Equation 1”. Based onthe disease incidences of the treated compartment and the untreatedcompartment, the control value of the treated compartment is calculatedby the following equation “Equation 2”, and the treated compartment isconfirmed to have a good control effect on plant disease.Disease incidence (%)=100×(number of diseased plant/total number ofseeded seeds):  Equation 1Control value (%)=100×[(disease incidence in untreated compartmentdisease incidence in treated compartment)/disease incidence in untreatedcompartment]:  Equation 2

The compartment treated with the composition of the invention shows asynergistic controlling effect, for each combination of the presentbacterial strain and the compound, compared with that of thecorresponding compartment treated solely with the present bacterialstrain or the compound.

TABLE 2 Retaining Bacteria/Compound Amount retained by Seeds (/Kg seeds)The Present Bacterial strain 1 × 10¹⁰ cfu The Present Bacterial strain 1× 10⁸ cfu The Present Bacterial strain 1 × 10⁶ cfu Ethaboxam 0.075 gEthaboxam 0.15 g Ethaboxam 0.3 g Metalaxyl 0.01 g Metalaxyl 0.04 gMetalaxyl 0.16 g The Present Bacterial strain 1 × 10¹⁰ cfu Ethaboxam0.075 g The Present Bacterial strain 1 × 10¹⁰ cfu Ethaboxam 0.15 g ThePresent Bacterial strain 1 × 10¹⁰ cfu Ethaboxam 0.3 g The PresentBacterial strain 1 × 10¹⁰ cfu Metalaxyl 0.01 g The Present Bacterialstrain 1 × 10¹⁰ cfu Metalaxyl 0.04 g The Present Bacterial strain 1 ×10¹⁰ cfu Metalaxyl 0.16 g The Present Bacterial strain 1 × 10⁸ cfuEthaboxam 0.3 g The Present Bacterial strain 1 × 10⁶ cfu Metalaxyl 0.01g The Present Bacterial strain 1 × 10⁸ cfu Metalaxyl 0.16 g

Test Example 17

In a rotary seed treatment machine (trade name: HEGE11, manufactured byWINTERSTEIGER), soybean seeds (variety: “Hatayutaka”) are treated bysmearing treatment using a chemical liquid, which has been prepared bydissolving a tolclofos-methyl flowable formulation (42% flowableformulation, trade name: Rizolex, Valent U.S.A. Corporation), afludioxonil flowable formulation (10% flowable formulation, trade name:Maxim100, Syngenta Crop Protection LLC) or the carboxamide compound (1)dissolved in acetone/Tween 20 (weight ratio=95:5) and diluted withwater, as well as wettable powder of the present bacterial strain asobtained in Formulation Example 11 and a flowable formulation of thepresent bacterial strain as obtained in Formulation Example 12, so thatthe soybean seeds retain the present bacterial strain and/or thecompound in the amount shown in Table 3.

In a rotary seed treatment machine (trade name: HEGE11, manufactured byWINTERSTEIGER), soybean seeds thus treated with tolclofos-methyl,fludioxonil or the carboxamide compound (1) are treated with wettablepowder of the present bacterial strain as obtained in FormulationExample 11 or a flowable formulation of the present bacterial strain asobtained in Formulation Example 12, so that the soybean seeds retain theamount shown in Table 3.

A plastic pot is filled with a soil, and then, the coon seeds, whichhave been treated with the present bacterial strain, compound or thepresent bacterial strain+compound as shown in Table 3, are seeded andcovered with a soil, which has been mixed with damping-off fungus(Rhizoctonia solani) cultured in a bran medium (“treated compartment”).Cultivation is carried out in a greenhouse under irrigation. The plantsare investigated 20 days after for the number of diseased plants, andthe disease incidence is calculated by the following “Equation 1”.Similar procedures are conducted using untreated corn seeds, instead ofthe treated corn seeds, as described above for the treated compartment(“untreated compartment”). The plants are investigated 20 days after forthe number of diseased plants, and the disease incidence is calculatedby the following “Equation 1”. Based on the disease incidences of thetreated compartment and the untreated compartment, the control value ofthe treated compartment is calculated by the following equation“Equation 2”, and the treated compartment is confirmed to have a goodcontrol effect on plant disease.Disease incidence (%)=100×(number of diseased plant/total number ofseeded seeds):  Equation 1Control value (%)=100×[(disease incidence in untreated compartmentdisease incidence in treated compartment)/disease incidence in untreatedcompartment]:  Equation 2

The compartment treated with the composition of the invention shows asynergistic controlling effect, for each combination of the presentbacterial strain and the compound, compared with that of thecorresponding compartment treated solely with the present bacterialstrain or the compound.

TABLE 3 Retaining Bacteria/Compound Amount retained by Seeds (/Kg seeds)The Present Bacterial strain 1 × 10¹⁰ cfu The Present Bacterial strain 1× 10⁶ cfu The Present Bacterial strain 1 × 10⁷ cfu The Present Bacterialstrain 1 × 10¹¹ cfu Tolclofos-Methyl 0.01 g Tolclofos-Methyl 0.05 gTolclofos-Methyl 0.25 g Fludioxonil 0.004 g Fludioxonil 0.02 gFludioxonil 0.1 g The Carboxamide Compound (1) 0.0004 g The CarboxamideCompound (1) 0.002 g The Carboxamide Compound (1) 0.01 g The PresentBacterial strain 1 × 10¹⁰ cfu Tolclofos-Methyl 0.01 g The PresentBacterial strain 1 × 10¹⁰ cfu Tolclofos-Methyl 0.05 g The PresentBacterial strain 1 × 10¹⁰ cfu Tolclofos-Methyl 0.25 g The PresentBacterial strain 1 × 10¹⁰ cfu Fludioxonil 0.004 g The Present Bacterialstrain 1 × 10¹⁰ cfu Fludioxonil 0.02 g The Present Bacterial strain 1 ×10¹⁰ cfu Fludioxonil 0.1 g The Present Bacterial strain 1 × 10¹⁰ cfu TheCarboxamide Compound (1) 0.0004 g The Present Bacteria 1 × 10¹⁰ cfu TheCarboxamide Compound (1) 0.002 g The Present Bacteria 1 × 10¹⁰ cfu TheCarboxamide Compound (1) 0.01 g The Present Bacterial strain 1 × 10⁶ cfuTolclofos-Methyl 0.01 g The Present Bacterial strain 1 × 10⁷ cfuTolclofos-Methyl 0.05 g The Present Bacterial strain 1 × 10⁶ cfuFludioxonil 0.004 g The Present Bacterial strain 1 × 10⁷ cfu Fludioxonil0.02 g The Present Bacterial strain 1 × 10⁷ cfu Fludioxonil 0.1 g ThePresent Bacterial strain 1 × 10¹¹ cfu The Carboxamide Compound (1)0.0004 g The Present Bacterial strain 1 × 10⁶ cfu The CarboxamideCompound (1) 0.01 g

Test Example 18

Wettable powder of the present bacterial strain as obtained inFormulation Example 11 or a flowable formulation of the presentbacterial strain as obtained in Formulation Example 12, ethaboxamflowable formulation (12.5% flowable formulation, trade name: Ethofinflowable, Nippon Soda Co., Ltd.) and metalaxyl are dissolved inacetone/Tween 20 (weight ratio=95:5) and diluted with water to prepare achemical solution, and the concentration is adjusted as shown in Table 4to prepare a spray liquid.

The liquid (50 mL) is sprayed to tomato plants (variety: Campari, 4-leafstage). After air drying, the plants are inoculated with late blightfungus (Phytophthora infestans) and left to stand under moisturecondition for 10 days (“treated compartment”). Also, similar proceduresare conducted without spraying the liquid (“untreated compartment”).

The effect on the treated compartment is determined by the followingequation, based on the onset area rates of the treated compartment andthe untreated compartment.controlling effect=100×[1−(onset area rate of treatedcompartment)/(onset area rate of untreated compartment)]  Equation:

The compartment treated with the composition of the invention shows asynergistic controlling effect, for each combination of the presentbacterial strain and the compound, compared with that of thecorresponding compartment treated solely with the present bacterialstrain or the compound.

TABLE 4 Bacteria/Compound Amount of Bacteria/Compound sprayed to Plantsin Spray Liquid (/L) The Present Bacterial strain 1 × 10⁸ cfu Ethaboxam0.4 mg Ethaboxam 0.8 mg Metalaxyl 1.5 mg Metalaxyl 3 mg The PresentBacterial strain 1 × 10⁸ cfu Ethaboxam 0.4 mg The Present Bacterialstrain 1 × 10⁸ cfu Ethaboxam 0.8 mg The Present Bacterial strain 1 × 10⁸cfu Metalaxyl 1.5 mg The Present Bacterial strain 1 × 10⁸ cfu Metalaxyl3 mg

Test Example 19

Wettable powder of the present bacterial strain as obtained inFormulation Example 11 or a flowable formulation of the presentbacterial strain as obtained in Formulation Example 12, fludioxonilflowable formulation (20% flowable formulation, trade name: Saviorflowable, Syngenta Japan) and the carboxamide compound (1) are dissolvedin acetone/Tween 20 (weight ratio=95:5) and diluted with water toprepare a chemical solution, and the concentration is adjusted as shownin Table 5 to prepare a spray liquid.

The liquid (50 mL) is sprayed to cucumber plants (variety: “SagamiHanjiro”, 2-leaf stage). After air drying, the plants are inoculatedwith gray mold fungus (Botrytis cinerea) and left to stand undermoisture condition for 10 days (“treated compartment”). Also, similarprocedures are conducted without spraying the liquid (“untreatedcompartment”).

The effect on the treated compartment is determined by the followingequation, based on the onset area rates of the treated compartment andthe untreated compartment.controlling effect=100×[1−(onset area rate of treatedcompartment)/(onset area rate of untreated compartment)]  Equation:

The compartment treated with the composition of the invention shows asynergistic controlling effect, for each combination of the presentbacterial strain and the compound, compared with that of thecorresponding compartment treated solely with the present bacterialstrain or the compound.

TABLE 5 Bacteria/Compound Amount of Bacteria/Compound sprayed to Plantsin Spray Liquid (/L) The Present Bacterial strain 1 × 10⁸ cfuFludioxonil 0.5 mg Fludioxonil 1 mg The Carboxamide Compound (1) 1 mgThe Carboxamide Compound (1) 4 mg The Present Bacterial strain 1 × 10⁸cfu Fludioxonil 0.5 mg The Present Bacterial strain 1 × 10⁸ cfuFludioxonil 1 mg The Present Bacterial strain 1 × 10⁸ cfu TheCarboxamide Compound (1) 1 mg The Present Bacterial strain 1 × 10⁸ cfuThe Carboxamide Compound (1) 4 mg

Test Example 20

A chemical liquid prepared by diluting tolclofos-methyl wettable powder(50% wettable powder, trade name: Rizolex 50, Sumitomo Chemical Co.,Ltd.) with water to 0.5 ppm or 2.5 ppm of tolclofos-methyl is combinedwith the equal volume of a solution of wettable powder of the presentbacterial strain as obtained in Formulation 11 or of a flowableformulation of the present bacterial strain as obtained in FormulationExample 12 (adjusted to 1×10⁸ cfu of the present bacterial strain per 1L of the liquid to be sprayed) (“treated compartment). Also, similarprocedures are conducted without spraying the liquid (“untreatedcompartment”).

The liquid (50 mL) is sprayed to sugar beet plants (variety: “Yukimaru”,growth stage). After air drying, the plants are inoculated with leafblight fungus (Thanatephorus cucumeris) and left to stand under moisturecondition for 10 days.

The effect on the treated compartment is determined by the followingequation, based on the onset area rates of the treated compartment andthe untreated compartment.controlling effect=100×[1−(onset area rate of treatedcompartment)/(onset area rate of untreated compartment)]  Equation:

The compartment treated with the composition of the invention shows asynergistic controlling effect, for each combination of the presentbacterial strain and the compound, compared with that of thecorresponding compartment treated solely with the present bacterialstrain or the compound.

Test Example 21

In a rotary seed treatment machine (trade name: HEGE11, manufactured byWINTERSTEIGER), corn seeds (variety: yellow dent corn) were treated bysmearing treatment using a flowable formulation of the present bacterialstrain as obtained in Formulation Example 13 and ethaboxam flowableformulation (34.6% flowable formulation, Valent U.S.A. Corporation), sothat the soybean seeds retain the present bacterial strain and/or thecompound in the amount shown in Table 6.

A plastic pot was filled with a soil, and then, the coon seeds, whichhave been treated with the present bacterial strain, compound or thepresent bacterial strain+compound as shown in Table 6, were seeded andcovered with a soil, which has been mixed with damping-off fungus(Pythium spp.) cultured in a bran medium or grass medium (“treatedcompartment”). Cultivation was carried out in a greenhouse underirrigation. Also, similar procedures were conducted using untreated cornseeds, instead of the treated corn seeds, as described above for thetreated compartment (“untreated compartment”). The plants wereinvestigated 20 days after for the number of diseased plants, and thedisease incidence was calculated by the following “Equation 1”. Thecontrol value of the treated compartment was calculated by the followingequation “Equation 2”, based on the disease incidences of the treatedcompartment and the untreated compartment.Disease incidence (%)=100×(number of diseased plant/total number ofseeded seeds):  Equation 1Control value (%)=100×[(disease incidence in untreated compartmentdisease incidence in treated compartment)/disease incidence in untreatedcompartment]:  Equation 2

TABLE 6 Retaining Bacteria/Compound Amount Control Estimated retained bySeeds (/kg seeds) Value Value* The Present Bacterial strain 1 × 10¹⁰ cfu9.7 — The Present Bacterial strain 1 × 10¹² cfu 12.9 — The PresentBacterial strain 1 × 10⁷ cfu 3.2 — Ethaboxam 0.00625 g 19.4 — Ethaboxam0.025 g 32.3 — Ethaboxam 0.1 g 48.4 — The Present Bacterial strain 1 ×10¹⁰ cfu 35.5 27.2 Ethaboxam 0.00625 g The Present Bacterial strain 1 ×10¹⁰ cfu 48.4 38.8 Ethaboxam 0.025 g The Present Bacterial strain 1 ×10¹⁰ cfu 77.4 53.4 Ethaboxam 0.1 g The Present Bacterial strain 1 × 10¹²cfu 51.6 29.8 Ethaboxam 0.00625 g The Present Bacterial strain 1 × 10¹²cfu 67.7 40.1 Ethaboxam 0.025 g The Present Bacterial strain 1 × 10⁷ cfu71.0 50.0 Ethaboxam 0.1 g *Control value estimated from the calculationby the Colby's equation

If the effect by the combination of two active ingredients is greaterthan that of the estimated value E, which is calculated by the Colby'sequation as follows, the effect is regarded as synergistic.E=X+Y−X·Y/100wherein,E=Control value when using the mixture of the active ingredients A and Bat the concentrations m and n (amount of the active ingredient),respectively.X=Control value when using the active ingredient A at the concentrationm (amount of the active ingredient).Y=Control value when using the active ingredient B at the concentrationn (amount of the active ingredient).

The compartment treated with the composition of the invention showed asynergistic controlling effect, for each combination of the presentbacterial strain and the compound, compared with that of thecorresponding compartment treated solely with the present bacterialstrain or the compound.

Test Example 22

Soybean seeds (variety:“Hatayutaka”) were treated by smearing treatmentusing a flowable formulation of the present bacterial strain as obtainedin Formulation Example 13, ethaboxam flowable formulation (34.6%flowable formulation, Valent U.S.A. Corporation) and metalaxyl flowableformulation (30.14% flowable formulation, trade name: Sebring, Nufarm),so that the soybean seeds retain the present bacterial strain and/or thecompound in the amount shown in Table 7.

A plastic pot was filled with a soil, and then, the soybean seeds, whichhave been treated with the present bacterial strain, compound or thepresent bacterial strain+compound as shown in Table 7, were seeded andcovered with a soil, which has been mixed with damping-off fungus(Pythium spp.) cultured in a bran medium or grass medium (“treatedcompartment”). Cultivation was carried out in a greenhouse underirrigation. Also, similar procedures were conducted using untreatedsoybean seeds, instead of the treated soybean seeds, as described abovefor the treated compartment (“untreated compartment”). The plants wereinvestigated 20 days after for the number of diseased plants, and thedisease incidence was calculated by the following “Equation 1”. Thecontrol value of the treated compartment was calculated by the followingequation “Equation 2”, based on the disease incidences of the treatedcompartment and the untreated compartment.Disease incidence (%)=100×(number of diseased plant/total number ofseeded seeds):  Equation 1Control value (%)=100×[(disease incidence in untreated compartmentdisease incidence in treated compartment)/disease incidence in untreatedcompartment]:  Equation 2

TABLE 7 Retaining Bacteria/Compound Amount Control Estimated retained bySeeds (/kg seeds) Value Value* The Present Bacterial strain 1 × 10¹⁰ cfu12.5 — The Present Bacterial strain 1 × 10⁸ cfu 9.4 — The PresentBacterial strain 1 × 10⁶ cfu 3.1 — Ethaboxam 0.075 g 46.9 — Ethaboxam0.15 g 56.2 — Ethaboxam 0.3 g 62.5 — Metalaxyl 0.01 g 40.6 — Metalaxyl0.04 g 50.0 — Metalaxyl 0.16 g 59.4 — The Present Bacterial strain 1 ×10¹⁰ cfu 65.6 53.5 Ethaboxam 0.075 g The Present Bacterial strain 1 ×10¹⁰ cfu 75.0 61.7 Ethaboxam 0.15 g The Present Bacterial strain 1 ×10¹⁰ cfu 84.4 67.2 Ethaboxam 0.3 g The Present Bacterial strain 1 × 10¹⁰cfu 62.5 48.0 Metalaxyl 0.01 g The Present Bacterial strain 1 × 10¹⁰ cfu75.0 56.3 Metalaxyl 0.04 g The Present Bacterial strain 1 × 10¹⁰ cfu81.3 64.5 Metalaxyl 0.16 g The Present Bacterial strain 1 × 10⁸ cfu 81.366.0 Ethaboxam 0.3 g The Present Bacterial strain 1 × 10⁶ cfu 53.1 42.4Metalaxyl 0.01 g The Present Bacterial strain 1 × 10⁸ cfu 78.1 63.2Metalaxyl 0.16 g *Control value estimated from the calculation by theColby's equation

The compartment treated with the composition of the invention showed asynergistic controlling effect, for each combination of the presentbacterial strain and the compound, compared with that of thecorresponding compartment treated solely with the present bacterialstrain or the compound.

Test Example 23

In a rotary seed treatment machine (trade name: HEGE11, manufactured byWINTERSTEIGER), soybean seeds (variety:“Hatayutaka”) were treated bysmearing treatment using a chemical liquid prepared by dissolvingTolclofos-methyl flowable formulation (42% flowable formulation, tradename: Rizolex, Valent U.S.A. Corporation), fludioxonil flowableformulation (20% flowable formulation, trade name: Savior flowable,Syngenta Japan) or the carboxamide compound (1) in acetone/Tween 20(weight ratio=95:5) and diluting with water, and a flowable formulationof the present bacterial strain as obtained in Formulation Example 13,so that the soybean seeds retain the present bacterial strain and/or thecompound in the amount shown in Table 8.

In a rotary seed treatment machine (trade name: HEGE11, manufactured byWINTERSTEIGER), soybean seeds thus treated with tolclofos-methyl,fludioxonil or the carboxamide compound (1) were treated with a flowableformulation of the present bacterial strain as obtained in FormulationExample 13, so that the soybean seeds retain the amount shown in Table8.

A plastic pot was filled with a soil, and then, the soybean seeds, whichhave been treated with the present bacterial strain, compound or thepresent bacterial strain+compound as shown in Table 8, were seeded andcovered with a soil, which has been mixed with damping-off fungus(Rhizoctonia solani) cultured in a bran medium. Cultivation was carriedout in a greenhouse under irrigation (“treated compartment”). Also,similar procedures were conducted using untreated soybean seeds, insteadof the treated soybean seeds, as described above for the treatedcompartment (“untreated compartment”). The plants were investigated 20days after for the number of diseased plants, and the disease incidencewas calculated by the following “Equation 1”. The control value of thetreated compartment was calculated by the following equation “Equation2”, based on the disease incidences of the treated compartment and theuntreated compartment.Disease incidence (%)=100×(number of diseased plant/total number ofseeded seeds):  Equation 1Control value (%)=100×[(disease incidence in untreated compartmentdisease incidence in treated compartment)/disease incidence in untreatedcompartment]:  Equation 2

TABLE 8 Retaining Bacteria/Compound Amount Control Estimated retained bySeeds (/kg seeds) Value Value* The Present Bacterial strain 1 × 10¹⁰ cfu14.2 — The Present Bacterial strain 1 × 10⁶ cfu 2.8 — The PresentBacterial strain 1 × 10⁷ cfu 5.7 — The Present Bacterial strain 1 × 10¹¹cfu 17.1 — Tolclofos-Methyl 0.01 g 11.4 — Tolclofos-Methyl 0.05 g 28.6 —Tolclofos-Methyl 0.25 g 42.8 — Fludioxonil 0.004 g 25.7 — Fludioxonil0.02 g 37.1 — Fludioxonil 0.1 g 48.6 — The Carboxamide Compound (1)0.0004 g 40.0 — The Carboxamide Compound (1) 0.002 g 54.3 — TheCarboxamide Compound (1) 0.01 g 65.7 — The Present Bacterial strain 1 ×10¹⁰ cfu 31.4 24.0 Tolclofos-Methyl 0.01 g The Present Bacterial strain1 × 10¹⁰ cfu 51.4 38.7 Tolclofos-Methyl 0.05 g The Present Bacterialstrain 1 × 10¹⁰ cfu 68.6 50.9 Tolclofos-Methyl 0.25 g The PresentBacterial strain 1 × 10¹⁰ cfu 48.6 36.3 Fludioxonil 0.004 g The PresentBacterial strain 1 × 10¹⁰ cfu 62.8 46.1 Fludioxonil 0.02 g The PresentBacterial strain 1 × 10¹⁰ cfu 74.3 55.9 Fludioxonil 0.1 g The PresentBacterial strain 1 × 10¹⁰ cfu 60.0 48.5 The Carboxamide Compound (1)0.0004 g The Present Bacterial strain 1 × 10¹⁰ cfu 77.1 60.8 TheCarboxamide Compound (1) 0.002 g The Present Bacterial strain 1 × 10¹⁰cfu 91.4 70.6 The Carboxamide Compound (1) 0.01 g The Present Bacterialstrain 1 × 10⁶ cfu 17.1 13.9 Tolclofos-Methyl 0.01 g The PresentBacterial strain 1 × 10⁷ cfu 42.8 32.6 Tolclofos-Methyl 0.05 g ThePresent Bacterial strain 1 × 10⁶ cfu 37.1 27.8 Fludioxonil 0.004 g ThePresent Bacterial strain 1 × 10⁷ cfu 54.3 40.7 Fludioxonil 0.02 g ThePresent Bacterial strain 1 × 10⁷ cfu 62.8 51.5 Fludioxonil 0.1 g ThePresent Bacterial strain 1 × 10¹¹ cfu 62.8 50.3 The Carboxamide Compound(1) 0.0004 g The Present Bacterial strain 1 × 10⁶ cfu 85.7 66.7 TheCarboxamide Compound (1) 0.01 g *Control value estimated from thecalculation by the Colby's equation

The compartment treated with the composition of the invention showed asynergistic controlling effect, for each combination of the presentbacterial strain and the compound, compared with that of thecorresponding compartment treated solely with the present bacterialstrain or the compound.

The invention claimed is:
 1. A composition for control plant diseasescomprising Bacillus strain APM-1 deposited under ATCC Accession No.PTA-4838 and one or more compounds selected from the group consisting ofethaboxam, tolclofos-methyl, metalaxyl, and fludioxonil, wherein thecomposition comprises 10⁻⁵ to 10² g of the compound per 10¹⁰ cfu ofBacillus strain APM-1.
 2. A method for controlling plant diseases,comprising a step of applying the composition defined in claim 1 to aplant or a plant cultivation site.
 3. The method for controlling plantdiseases according to claim 2 wherein the plant is a geneticallymodified plant.
 4. A plant seed or vegetative propagation organcomprising 10⁶ to 10¹³ cfu of Bacillus strain APM-1 and 0.0001 to 10 gof one or more compounds selected from the group consisting ofethaboxam, tolclofos-methyl, metalaxyl, and fludioxonil per 1 kg of theseed or vegetative propagation organ.